Celery cultivar tbg 45

ABSTRACT

A celery cultivar designated TBG 45 is disclosed. The invention relates to the seeds of celery cultivar TBG 45, to the plants of celery cultivar TBG 45 and to methods for producing a celery plant by crossing the cultivar TBG 45 with itself or another celery cultivar. The invention further relates to methods for producing a celery plant containing in its genetic material one or more transgenes and to the transgenic celery plants and plant parts produced by those methods. This invention also relates to celery cultivars or breeding cultivars and plant parts derived from celery cultivar TBG 45, to methods for producing other celery cultivars, lines or plant parts derived from celery cultivar TBG 45 and to the celery plants, varieties, and their parts derived from the use of those methods. The invention further relates to hybrid celery seeds, plants, and plant parts produced by crossing cultivar TBG 45 with another celery cultivar.

BACKGROUND OF THE INVENTION

The present invention relates to a new and distinctive celery (Apiumgraveolens var. dulce) variety designated TBG 45. All publications citedin this application are herein incorporated by reference.

There are numerous steps in the development of any novel, desirableplant germplasm. Plant breeding begins with the analysis, definition ofproblems and weaknesses of the current germplasm, the establishment ofprogram goals, and the definition of specific breeding objectives. Thenext step is selection of germplasm that possesses the traits to meetthe program goals. The goal is to combine in a single variety or hybridan improved combination of desirable traits from the parental germplasm.These important traits may include improved flavor, increased stalk sizeand weight, higher seed yield, improved color, resistance to diseasesand insects, tolerance to drought and heat, and better agronomicquality.

All cultivated forms of celery belong to the species Apium graveolensvar. dulce that is grown for its edible stalk. As a crop, celery isgrown commercially wherever environmental conditions permit theproduction of an economically viable yield. In the United States, theprincipal growing regions are California, Florida, Arizona, andMichigan. Fresh celery is available in the United States year-round,although the greatest supply is from November through January. Forplanting purposes, the celery season is typically divided into twoseasons: summer and winter, with Florida and the southern Californiaareas harvesting from November to July, and Michigan and northernCalifornia harvesting from July to October. Celery is consumed as fresh,raw product and as a cooked vegetable.

Celery is a cool-season biennial that grows best from 60° F. to 65° F.(16° C. to 18° C.) but will tolerate temperatures from 45° F. to 75° F.(7° C. to 24° C.). Freezing damages mature celery by splitting thepetioles or causing the skin to peel, making the stalks unmarketable.This can be a problem for crops planted in the winter regions; however,celery can tolerate minor freezes early in the production cycle.

The two main growing regions for celery in California are located alongthe Pacific Ocean: the central coast or summer production area(Monterey, San Benito, Santa Cruz, and San Luis Obispo Counties) and thesouth coast or winter production area (Ventura and Santa BarbaraCounties). A minor region (winter) is located in the southern deserts(Riverside and Imperial Counties).

In the south coast, celery is transplanted from early August to Aprilfor harvest from November to mid-July; in the Santa Maria area, celeryis transplanted from January to August for harvest from April throughDecember. In the central coast, fields are transplanted from March toSeptember for harvest from late June to late December. In the southerndeserts, fields are transplanted in late August for harvest in January.

Commonly used celery varieties for coastal production include Command,Mission, Conquistador and Sonora. Some shippers use their ownproprietary varieties. Celery seed is very small and difficult togerminate. All commercial celery is planted as greenhouse-growntransplants. Celery grown from transplants is more uniform than fromseed and takes less time to grow the crop in the field. Transplantedcelery is traditionally placed in double rows on 40-inch (100-cm) bedswith plants spaced between 6.0 and 7.0 inches apart.

Celery requires a relatively long and cool growing season (Thephysiology of vegetable crops by Pressman, CAB Intl., New York, 1997).Earlier transplanting results in a longer growing season, increasedyields, and better prices. However, celery scheduled for Spring harvestoften involves production in the coolest weather conditions of Winter, aperiod during which vernalization can occur. If adequate vernalizationoccurs for the variety, bolting may be initiated. Bolting is thepremature rapid elongation of the main celery stem into a floral axis(i.e., during the first year for this normally biennial species).Bolting slows growth as the plant approaches marketable size and leavesa stalk with no commercial value. Different varieties have differentvernalization requirements, but in the presence of bolting, the lengthof the seed stem can be used as a means of measuring bolting tolerancethat exists in each different variety. The most susceptible varietiesreach their vernalization requirement earlier and have time to developthe longest seed stems, while the moderately tolerant varieties takelonger to reach their vernalization requirement and have less time todevelop a seed stem which would therefore be shorter. Under normalproduction conditions, the most tolerant varieties may not achieve theirvernalization requirement and therefore not produce a measurable seedstem.

The coldest months when celery is grown in the United States areDecember, January and February. If celery is going to reach itsvernalization requirements to cause bolting, it is generally youngercelery that is exposed to this cold weather window. This celerygenerally matures in the months of April and May which constitute whatthe celery industry calls the bolting or seeder window. The bolting orseeder window is a period where seed stems are generally going to impactthe quality of the marketable celery, and this is most consistentlyexperienced in celery grown in the Southern California region. Thepresence of seed stems in celery can be considered a major marketabledefect as set forth in the USDA grade standards. If the seed stem islonger than twice the diameter of the celery stalk or eight inches, thecelery no longer meets the standards of US Grade #1. If the seed stem islonger than three times the diameter of the celery stalk, the celery isno longer marketable as US Grade #2 (United States Standards for Gradesof Celery, United States Department of Agriculture, reprinted January1997).

Celery is an allogamous biennial crop. The celery genome consists of 11chromosomes. Its high degree of out-crossing is accomplished by insectsand wind pollination. Pollinators of celery flowers include a largenumber of wasp, bee and fly species. Celery is subject to inbreedingdepression, which appears to be dependent upon the genetic background assome lines are able to withstand selfing for three or four generations.

Celery flowers are protandrous, with pollen being released 3-6 daysbefore stigma receptivity. At the time of stigma receptivity, thestamens will have fallen, and the two stigmata will have unfolded in anupright position. The degree of protandry varies, which makes itdifficult to perform reliable hybridization, due to the possibility ofaccidental selfing.

Celery flowers are very small, which significantly hinders easy removalof individual anthers. Furthermore, different developmental stages ofthe flowers in umbels make it difficult to avoid uncontrolledpollinations. The standard hybridization technique in celery consists ofselecting flower buds of the same size and eliminating the older andyounger flowers. Then, the umbellets are covered with glycine paper bagsfor a 5-10 day period, during which the stigmas become receptive. At thetime the flowers are receptive, available pollen or umbellets sheddingpollen from selected male parents are rubbed on to the stigmas of thefemale parent.

Celery plants require a period of vernalization while in the vegetativephase in order to induce seed stalk development. A period of 6-10 weeksat 5° C. to 8° C. when the plants are greater than 4 weeks old isusually adequate for most non-bolting tolerant varieties. Due to a widerange of responses to the cold treatment, it is often difficult tosynchronize crossing, since plants will flower at different times.However, pollen can be stored for 6-8 months at −10° C. in the presenceof silica gel or calcium chloride with a viability decline of only20-40%, thus providing flexibility to perform crosses over a longertime.

For selfing, the plant or selected umbels are caged in cloth bags. Theseare shaken several times during the day to promote pollen release.Houseflies (Musca domestica) can also be introduced weekly into the bagsto perform pollinations.

Celery in general is an important and valuable vegetable crop. Thus, acontinuing goal of celery plant breeders is to develop stable, highyielding celery cultivars that are resistant to diseases andagronomically sound to maximize the amount of yield produced on theland. To accomplish this goal, the celery breeder must select anddevelop celery plants that have the traits that result in superiorcultivars.

The foregoing examples of the related art and limitations relatedtherewith are intended to be illustrative and not exclusive. Otherlimitations of the related art will become apparent to those of skill inthe art upon a reading of the specification.

SUMMARY OF THE INVENTION

The following embodiments and aspects thereof are described inconjunction with systems, tools, and methods which are meant to beexemplary and illustrative, not limiting in scope. In variousembodiments, one or more of the above-described problems have beenreduced or eliminated, while other embodiments are directed to otherimprovements.

According to the invention, there is provided a novel celery cultivardesignated TBG 45. Also provided are celery plants having thephysiological and morphological characteristics of celery cultivar TBG45. This invention thus relates to the seeds of celery cultivar TBG 45,to the plants of celery cultivar TBG 45, and to methods for producing acelery plant by crossing celery TBG 45 with itself or another celeryplant, to methods for producing a celery plant containing in its geneticmaterial one or more transgenes and to the transgenic celery plantsproduced by that method. This invention also relates to methods forproducing other celery cultivars derived from celery cultivar TBG 45 andto the celery cultivar derived by the use of those methods. Thisinvention further relates to hybrid celery seeds and plants produced bycrossing celery cultivar TBG 45 with another celery line.

This invention further relates to the F₁ hybrid celery plants and plantparts grown from the hybrid seed produced by crossing celery cultivarTBG 45 to a second celery plant. Still further included in the inventionare the seeds of an F₁ hybrid plant produced with the celery cultivarTBG 45 as one parent, the second generation (F₂) hybrid celery plantgrown from the seed of the F₁ hybrid plant, and the seeds of the F₂hybrid plant. Thus, any such methods using the celery cultivar TBG 45are part of this invention: selfing, backcrosses, hybrid production,crosses to populations, and the like. All plants produced using celerycultivar TBG 45 as at least one parent are within the scope of thisinvention. Advantageously, the celery cultivar could be used in crosseswith other, different, celery plants to produce first generation (F₁)celery hybrid seeds and plants with superior characteristics.

The invention further provides methods for developing celery plantsderived from celery cultivar TBG 45 in a celery plant breeding programusing plant breeding techniques including recurrent selection,backcrossing, pedigree breeding, restriction fragment lengthpolymorphism enhanced selection, genetic marker enhanced selection andtransformation. Seeds, celery plants, and parts thereof, produced bysuch breeding methods are also part of the invention.

In another aspect, the present invention provides protoplasts andregenerable cells for use in tissue culture of celery cultivar TBG 45.The tissue culture will preferably be capable of regenerating plantshaving essentially all of the physiological and morphologicalcharacteristics of the foregoing celery plant, and of regeneratingplants having substantially the same genotype as the foregoing celeryplant. Preferably, the regenerable cells in such tissue cultures will becallus, protoplasts, meristematic cells, leaves, pollen, embryos, roots,root tips, anthers, pistils, flowers, seeds, petioles and suckers. Stillfurther, the present invention provides celery plants regenerated fromthe tissue cultures of the invention.

In another aspect, the present invention provides for single or multiplegene converted plants of TBG 45. The single or multiple transferredgene(s) may preferably be a dominant or recessive allele. Preferably,the single or multiple transferred gene(s) will confer such traits asmale sterility, herbicide resistance, insect resistance, modified fattyacid metabolism, modified carbohydrate metabolism, resistance forbacterial, fungal, or viral disease, male fertility, enhancednutritional quality and industrial usage or the transferred gene willhave no apparent value except for the purpose of being a marker forvariety identification. The single or multiple gene(s) may be anaturally occurring celery gene or a transgene introduced throughgenetic engineering techniques.

The invention also relates to methods for producing a celery plantcontaining in its genetic material one or more transgenes and to thetransgenic celery plant produced by those methods.

In another aspect, the present invention provides for methods ofintroducing one or more desired trait(s) into celery cultivar TBG 45 andplants or seeds obtained from such methods. The desired trait(s) may be,but not exclusively, a single gene, preferably a dominant but also arecessive allele. Preferably, the transferred gene or genes will confersuch traits as male sterility, herbicide resistance, insect resistance,disease resistance, resistance for bacterial, fungal, or viral disease,male fertility, water stress tolerance, enhanced nutritional quality,modified fatty acid metabolism, modified carbohydrate metabolism,enhanced plant quality, and industrial usage. The gene or genes may benaturally occurring rice gene(s). The method for introducing the desiredtrait(s) may be a backcrossing process making use of a series ofbackcrosses to the celery cultivar TBG 45 during which the desiredtrait(s) is maintained by selection. The desired trait may also beintroduced via transformation.

The invention further relates to methods for genetically modifying acelery plant of the celery cultivar TBG 45 and to the modified celeryplant produced by those methods. The genetic modification methods mayinclude, but are not limited to mutation, genome editing, genesilencing, RNA interference, backcross conversion, genetictransformation, single and multiple gene conversion, and/or direct genetransfer. The invention further relates to a genetically modified celeryplant produced by the above methods, wherein the genetically modifiedcelery plant comprises the genetic modification and otherwise comprisesall of the physiological and morphological characteristics of celerycultivar TBG 45.

In addition to the exemplary aspects and embodiments described above,further aspects and embodiments will become apparent by reference by thestudy of the following descriptions.

DETAILED DESCRIPTION OF THE INVENTION

In the description and tables which follow, a number of terms are used.In order to provide a clear and consistent understanding of thespecification and claims, including the scope to be given such terms,the following definitions are provided:

Allele. An allele is any of one or more alternative form of a gene, allof which relate to one trait or characteristic. In a diploid cell ororganism, the two alleles of a given gene occupy corresponding loci on apair of homologous chromosomes.

Backcrossing. Backcrossing is a process in which a breeder repeatedlycrosses hybrid progeny back to one of the parents, for example, a firstgeneration hybrid F₁ with one of the parental genotype of the F₁ hybrid.

Bacterial blight. A bacterial disease of celery caused by Pseudomonassyringae pv. apii. The initial symptoms appear on the leaves as small,bright yellow, circular spots. As these enlarge with a yellow halo, theyturn to a rust color. As the spots increase in number they merge toeventually kill the leaf tissue. Bacterial blight is favored by cool,wet conditions and at least 10 hours of leaf wetness is required forinfection. The disease is spread by water splashes, farm machinery andfield workers especially when the foliage is wet.

Black streak. A physiological disorder in celery plants causing somepetioles, when cut, to show “black streaks” in the lower half orthroughout the entire length of the petiole, making the entire cropunmarketable. Symptoms can be triggered under field conditions by hightemperatures.

Blackheart. Blackheart is due to a lack of movement of sufficientcalcium that causes the plant to turn brown and begin to decay at thegrowing point of the plant. Celery in certain conditions, such as warmweather, grows very rapidly and is incapable of moving sufficientamounts of calcium to the growing point.

Bolting. The premature development of a flowering or seed stalk, andsubsequent seed, before a plant produces a food crop. Bolting istypically caused by late planting when temperatures are low enough tocause vernalization of the plants.

Bolting period. Also known as the bolting or seeder window, andgenerally occurs in celery that is transplanted from the middle ofDecember through January and matures in April to May. The intensity andactual weeks that bolting may be observed vary from year to year, but itis generally observed in this window.

Bolting tolerance. The amount of vernalization that is required fordifferent celery varieties to bolt is genetically controlled. Varietieswith increased tolerance to bolting require greater periods ofvernalization in order to initiate bolting. A comparison of boltingtolerance between varieties can be measured by the length of theflowering or seed stem under similar vernalization conditions.

Brown stem. A disease caused by the bacterium Pseudomonas cichorii thatcauses petiole necrosis. Brown Stem is characterized by a firm, browndiscoloration throughout the petiole.

Celeriac or Root celery (Apium graveolens L. var. rapaceum). A plantthat is related to celery but instead of having a thickened andsucculent leaf petiole as in celery, celeriac has an enlarged hypocotyland upper root that is the edible product.

Celery heart. The center most interior petioles and leaves of the celerystalk. They are not only the smallest petioles in the stalk, but theyoungest as well. Some varieties are considered heartless because theygo right from very large petioles to only a couple of very smallpetioles. The heart is comprised of the petioles that are closest to themeristem of the celery stalk.

Colletotrichum. One of the most common and important genera ofplant-pathogenic fungi. Causes post-harvest rots, and anthracnose spotsand blights of aerial plant parts. In celery it is also frequentlyaccompanied by curling of the foliage and black heart. Colletotrichumacutatum sensu lato has been reclassified to be Colletotrichum fioriniae(Pavel, J. A., The Etiology, Virulence, and Phylogenetics of the CeleryAnthracnose Pathogen, Colletotrichum fioriniae (=C. acutatum sensulato), Graduate Theses and Dissertations (2016)).

Consumable. Means material that is edible by humans.

Crackstem. The petiole can crack or split horizontally orlongitudinally. Numerous cracks in several locations along the petioleare often an indication that the variety has insufficient boronnutrition. A variety's ability to utilize boron is a physiologicalcharacteristic which is genetically controlled.

Essentially all of the physiological and morphological characteristics.A plant having essentially all of the physiological and morphologicalcharacteristics of a designated plant has all of the characteristics ofthe plant that are otherwise present when compared in the sameenvironment, other than an occasional variant trait that might ariseduring backcrossing or direct introduction of a transgene.

F_(#). The “F” symbol denotes the filial generation, and the # is thegeneration number, such as F₁, F₂, F₃, etc.

F₁ hybrid. The first generation progeny of the cross of two nonisogenicplants.

Feather leaf. Feather Leaf is a yellowing of the lower leaflets andgenerally occurs in the outer petioles but can also be found on innerpetioles of the stalk. These yellowing leaves which would normallyremain in the harvested stalk are considered unacceptable. Thesepetioles then have to be stripped off in order to meet USDA standardswhich effectively decreases the stalk size and yield.

Flare. The lower, generally wider portion of the petiole which isusually a paler green or white. Some also refer to this as the spoon,scoop, or shovel.

Fusarium yellows. A fungal soilborne disease caused by Fusariumoxysporum f. sp. apii Race 2 and/or Race 4. Infected plants turn yellowand are stunted. Some of the large roots may have a dark brown and awater-soaked appearance. The water-conducting tissue (xylem) in thestem, crown, and root show a characteristic orange-brown discoloration.In the later stages of infection, plants remain severely stunted andyellowed and may collapse. The disease appears most severe during warmseasons, and in heavy, wet soils. Race 4 may also produce symptoms ofblack heart.

Gene. As used herein, “gene” refers to a segment of nucleic acid. A genecan be introduced into a genome of a species, whether from a differentspecies or from the same species, using transformation or variousbreeding techniques.

Gene silencing. The interruption or suppression of the expression of agene at the level of transcription or translation.

Genetically modified. Describes an organism that has received geneticmaterial from another organism, or had its genetic material modified,resulting in a change in one or more of its phenotypic characteristics.Methods used to modify, introduce or delete the genetic material mayinclude mutation breeding, genome editing, RNA interference, backcrossconversion, genetic transformation, single and multiple gene conversion,and/or direct gene transfer.

Genome editing. A type of genetic engineering in which DNA is inserted,replaced, modified or removed from a genome using artificiallyengineered nucleases or other targeted changes using homologousrecombination. Examples include but are not limited to use of zincfinger nucleases (ZFNs), TAL effector nucleases (TALENs), endonucleases,meganucleases, CRISPR/Cas9, and other CRISPR related technologies. (Maet. al., Molecular Plant, 9:961-974 (2016); Belhaj et. al., CurrentOpinion in Biotechnology, 32:76-84 (2015)).

Gross yield (Pounds/Acre). The total yield in pounds/acre of trimmedcelery plants (stalks).

Leaf celery (Apium graveolens L. var. secalinum). A plant that has beendeveloped primarily for leaf and seed production. Often grown inMediterranean climates, leaf celery more closely resembles celery's wildancestors. The stems are small and fragile and vary from solid to hollowand the leaves are fairly small and are generally bitter. This type isoften used for its medicinal properties and spice.

Leaf margin chlorosis. A magnesium deficiency producing an interveinalchlorosis which starts at the margin of leaves.

Maturity date. Maturity in celery can be dictated by two conditions. Thefirst, or true maturity, is the point in time when the celery reachesmaximum size distribution, but before defects such as pith, yellowing,Feather Leaf or Brown Stem appear. The second, or market maturity is anartificial maturity dictated by market conditions, i.e, the marketrequirement may be for 3 dozen sizes so the field is harvested atslightly below maximum yield potential because the smaller sizes arewhat the customers prefer at that moment.

Muck. Muck is a soil made up primarily of humus drained from swampland.It is used for growing specialty crops, such as onions, carrots, celery,and potatoes.

MUN. MUN refers to the MUNSELL Color Chart which publishes an officialcolor chart for plant tissues according to a defined numbering system.The chart may be purchased from the Macbeth Division of KollmorgenInstruments Corporation, 617 Little Britain Road, New Windsor, New York12553-6148.

Petiole. A petiole is the stem or limb of a leaf, the primary portion ofthe celery consumed.

Petiole depth. The average measurement in millimeters of the depth ofthe celery petiole at its narrowest point. The petiole depth measurementis taken from the outside of the petiole (which is the part of thepetiole that faces the outside of the stalk) and is measured to theinside of the petiole or cup or the inner most point of the petiole thatfaces the center of the stalk or heart.

Petiole width. The average measurement of the width of the celerypetiole in millimeters at its widest point. The measurement is takenfrom the side or edge of petiole to the opposite side or edge of thepetiole. The measurement is taken 90 degrees from petiole depth.

Phthalides. One of the chemical compounds that are responsible for thecharacteristic flavor and aroma of celery.

Pith. Pith is a sponginess/hollowness/white discoloration that occurs inthe petioles of celery varieties naturally as they become over-mature.In some varieties it occurs at an earlier stage causing harvest to occurprior to ideal maturity. Pith generally occurs in the outer, olderpetioles first. If it occurs, these petioles are stripped off to makegrade, which effectively decreases the stalk size and overall yieldpotential.

Plant height. The height of the plant from the bottom of the base orbutt of the celery plant to the top of the tallest leaf.

Polyphenol oxidase (PPO). An enzyme that catalyzes the conversion ofphenolic compounds to quinones and assists their products'polymerization. The catalysis of PPO, in the presence of oxygen, leadsto the formation of undesirable brown pigments and off-flavoredproducts.

Quantitative Trait Loci. Quantitative Trait Loci (QTL) refers to geneticloci that control to some degree, numerically representable traits thatare usually continuously distributed.

Regeneration. Regeneration refers to the development of a plant fromtissue culture.

Ribbing. The texture of the exterior surface of the celery petiole canvary from smooth to ribby depending on the variety. Ribbing is thepresence of numerous ridges that run vertically along the petioles ofthe celery plant.

Seed stem. A seed stem is the result of the elongation of the main stemof the celery, which is usually very compressed to almost non-existent,to form the flowering axis. The seed stem or flowering axis can reachseveral feet in height during full flower. The length of the seed stemis measured as the distance from the top of the basal plate (the base ofthe seed stem) to its terminus (the terminal growing point).

Septoria apiicola. A fungus that is the cause of late blight in celery.Symptoms include chlorotic spots that turn brown and necrotic.

Single gene converted. Single gene converted or conversion plant refersto plants which are developed by backcrossing, or via geneticengineering, wherein essentially all of the desired morphological andphysiological characteristics of a line are recovered in addition to thesingle gene transferred into the line via the backcrossing technique orvia genetic engineering.

Stalk. A stalk is a single celery plant that is trimmed with the top orfoliage and the roots removed.

Standard stem celery. A more traditional stem celery with moderate jointlength, to be utilized and marketed as a whole stalk with 12 to 14 inchcut or for hearts in retail environment.

Stringiness. Stringiness is a physiological characteristic that isgenerally associated with strings that get stuck between the consumer'steeth. There are generally two sources of strings in celery. One is thevascular bundle which can be fairly elastic and behave as a string. Thesecond is a strip of particularly strong epidermis cells calledschlerenchyma which are located on the surface of the ridges of thecelery varieties that have ribs.

Suckers. Suckers are auxiliary shoots that form at the base of the stalkor within the auxiliary buds between each petiole. If these shoots formbetween the petioles of the stalk, several petioles have to be strippedoff causing the celery to become smaller and the functional yields to bedecreased.

Tall stem celery. A stem celery with especially long petioles withprimary purpose of being utilized for production of sticks or limbs.

Transgene. A nucleic acid of interest that can be introduced into thegenome of a plant by genetic engineering techniques (e.g.,transformation) or breeding.

Celery cultivar TBG 45 is a new traditional carton-style celery cultivarthat has shown tremendous potential for production in the Ventura Countyproduction district of California. It has been developed by recurringsingle plant selection under conditions with very high Fusariumoxysporum f. sp. apii race 2 followed by selection for freeness fromFusarium oxysporum f. sp. apii race 4 infections. As a result, celerycultivar TBG 45 has excellent tolerance to race 2 and very goodtolerance to race 4. This combination of tolerance capabilities allowsTBG 45 to perform better than other varieties in Ventura County ofCalifornia where many fields are transitioning from race 2 infection toboth race 2 and race 4. Since there is not currently an adequate meansto determine inoculum levels of the different fusarium in the soil,hence infectious capabilities, both of which are very dependent onclimatic conditions, it is not possible to adequately ensure yields withcultivars that have insufficient fusarium tolerance for either race, sohaving tolerance to race 4 in combination with excellent tolerance torace 2 is extremely important.

In fields where there is a known presence of both race 2 and higherinfectious level for race 4, celery cultivar TBG 45 is the bestcandidate for performance. In the presence of race 2 only, TBG 29remains difficult to outperform based on yield potential; however, withlow to high race 4 pressure TBG 45 fairly consistently out yields TBG 29along with most other cultivars. With higher infection rates for race 4,TBG 45 outperforms TBG 43.

Celery cultivar TBG 45 has slight tolerance to bolting, so it hasviability for production in the late spring (June-July) when there canstill be slight residual winter bolting pressure combined with warmweather conditions that promote infection from both fusarium race 2 and4.

Celery cultivar TBG 45 also performs very well at higher plantpopulations and has good tolerance to most defects which can become veryproblematic for most celery varieties at higher plant populations.However, like most varieties care must be taken to time harvestappropriately in order to ensure that pith does not become a majordefect that occurs with over maturity.

Feather leaf and suckers are also slightly higher risk and like mostvarieties that require monitoring for pith as a gauge for timingmaturity and corresponding harvest, feather leaf may be the gauge forTBG 45. Suckers appear to be more of a spring issue when boltingpressure appears to coincide with them.

Celery cultivar TBG 45 has the following morphologic and othercharacteristics (based primarily on data collected in California):

Table 1 Variety Description Information

Maturity: 100 days in Oxnard, California

Plant height: 81.7 cm

Number of outer petioles (>35.6 cm): 11.95

Number of inner petioles (<35.6 cm): 4.75

Stalk shape: Cylindrical

Stalk conformation: Fairly compact

Heart formation: Full heart

Petiole length (from butt to first joint): 30.8 cm

Petiole width (at petiole midpoint): 24.4 mm

Petiole thickness (at petiole midpoint): 11.4 mm

Petiole cross section shape: Slight cup to cup

Petiole color (un-blanched at harvest): MUN 5gy 6/6

Anthocyanin: Absent

Stringiness: Very slight

Ribbing: Smooth to slight rib

Glossiness: Glossy

Leaf blade color: MUN 5gy 3/4

Bolting tolerance: Slight

Stress tolerance:

-   -   Adaxial crackstem (boron deficiency): Tolerant    -   Abaxial crackstem (boron deficiency): Tolerant    -   Leaf margin chlorosis (magnesium deficiency): Tolerant    -   Blackheart (calcium deficiency): Tolerant    -   Pithiness (nutritional deficiency): Moderately tolerant    -   Feather leaf: Moderately tolerant    -   Sucker development: Moderately tolerant

Disease resistance:

-   -   Brown stem (Pseudomonas cichorii): Tolerant    -   Bacterial blight (Pseudomonas syringae pv. apii): Unknown    -   Late blight (Septoria apii): Susceptible    -   Fusarium oxysporum f. sp. apii race 2: Very tolerant    -   Fusarium oxysporum f. sp. apii race 4: Tolerant

This invention is also directed to methods for producing a celery plantby crossing a first parent celery plant with a second parent celeryplant, wherein the first parent celery plant or second parent celeryplant is celery cultivar TBG 45. Further, both the first parent celeryplant and second parent celery plant may be from celery cultivar TBG 45.Still further, this invention also is directed to methods for producinga cultivar TBG 45-derived celery plant by crossing cultivar TBG 45 witha second celery plant and growing the progeny seed, and repeating thecrossing and growing steps with the cultivar TBG 45-derived plant from 0to 7 times. Thus, any such methods using the cultivar TBG 45 are part ofthis invention: selfing, backcrosses, hybrid production, crosses topopulations, and the like. All plants produced using cultivar TBG 45 asa parent are within the scope of this invention, including plantsderived from cultivar TBG 45. Advantageously, cultivar TBG 45 can beused in crosses with other, different cultivars to produce firstgeneration (F₁) celery seeds and plants with superior characteristics.

Additional methods include, but are not limited to, expression vectorsintroduced into plant tissues using a direct gene transfer method suchas microprojectile-mediated delivery, DNA injection, electroporation andthe like. More preferably, expression vectors are introduced into planttissues by using either microprojectile-mediated delivery with abiolistic device or by using Agrobacterium-mediated transformation.Transformant plants obtained with the protoplasm of the invention areintended to be within the scope of this invention.

As used herein, the term “plant” includes plant cells, plantprotoplasts, plant cell tissue cultures from which celery plants can beregenerated, plant calli, plant clumps and plant cells that are intactin plants or parts of plants, such as leaves, pollen, embryos,meristematic cells, hypocotyls, roots, root tips, anthers, pistils,flowers, seeds, stems, and the like.

Further Embodiments of the Invention

Celery in general is an important and valuable vegetable crop. Thus, acontinuing goal of celery plant breeders is to develop stable, highyielding celery cultivars that are agronomically sound. To accomplishthis goal, the celery breeder must select and develop celery plants withtraits that result in superior cultivars.

Plant breeding techniques known in the art and used in a celery plantbreeding program include, but are not limited to, pedigree breeding,recurrent selection, mass selection, single or multiple-seed descent,bulk selection, backcrossing, open pollination breeding, restrictionfragment length polymorphism enhanced selection, genetic marker enhancedselection, making double haploids, and transformation. Oftencombinations of these techniques are used. The development of celeryvarieties in a plant breeding program requires, in general, thedevelopment and evaluation of homozygous varieties. There are manyanalytical methods available to evaluate a new variety. The oldest andmost traditional method of analysis is the observation of phenotypictraits, but genotypic analysis may also be used.

Using Celery Cultivar TBG 45 to Develop Other Celery Varieties

This invention also is directed to methods for producing a celery plantby crossing a first parent celery plant with a second parent celeryplant wherein the first or second parent celery plant is a celery plantof cultivar TBG 45. Further, both first and second parent celery plantscan come from celery cultivar TBG 45. Also provided are methods forproducing a celery plant having substantially all of the morphologicaland physiological characteristics of cultivar TBG 45, by crossing afirst parent celery plant with a second parent celery plant wherein thefirst and/or the second parent celery plant is a plant havingsubstantially all of the morphological and physiological characteristicsof cultivar TBG 45 as determined at the 5% significance level when grownin the same environmental conditions. The other parent may be any celeryplant, such as a celery plant that is part of a synthetic or naturalpopulation. Thus, any such methods using celery cultivar TBG 45 are partof this invention: selfing, backcrosses, hybrid production, crosses topopulations, and the like. All plants produced using celery cultivar TBG45 as at least one parent are within the scope of this invention,including those developed from cultivars derived from celery cultivarTBG 45.

The cultivar of the invention can also be used for transformation whereexogenous genes are introduced and expressed by the cultivar of theinvention. Genetic variants created either through traditional breedingmethods using celery cultivar TBG 45 or through transformation ofcultivar TBG 45 by any of a number of protocols known to those of skillin the art are intended to be within the scope of this invention.

The following describes breeding methods that may be used with celerycultivar TBG 45 in the development of further celery plants. One suchembodiment is a method for developing a progeny celery plant in a celeryplant breeding program comprising: obtaining the celery plant, or a partthereof, of cultivar TBG 45, utilizing said plant or plant part as asource of breeding material, and selecting a celery cultivar TBG 45progeny plant with molecular markers in common with cultivar TBG 45and/or with morphological and/or physiological characteristics of celerycultivar TBG 45. Breeding steps that may be used in the celery plantbreeding program include, but are not limited to, pedigree breeding,backcrossing, mutation breeding, and recurrent selection. In conjunctionwith these steps, techniques such as RFLP-enhanced selection, geneticmarker enhanced selection (for example SSR markers) and the making ofdouble haploids may be utilized.

Another method involves producing a population of celery cultivar TBG 45progeny celery plants, comprising crossing cultivar TBG 45 with anothercelery plant, thereby producing a population of celery plants, which, onaverage, derive 50% of their alleles from celery cultivar TBG 45. Aplant of this population may be selected and repeatedly selfed or sibbedwith a celery cultivar resulting from these successive filialgenerations. One embodiment of this invention is the celery cultivarproduced by this method and that has obtained at least 50% of itsalleles from celery cultivar TBG 45.

Still yet another aspect of the invention is a method of producing acelery plant derived from the celery cultivar TBG 45, the methodcomprising the steps of: (a) preparing a progeny plant derived fromcelery cultivar TBG 45 by crossing a plant of the celery cultivar TBG 45with a second celery plant; and (b) crossing the progeny plant withitself or a second plant to produce a seed of a progeny plant of asubsequent generation which is derived from a plant of the celerycultivar TBG 45. In further embodiments of the invention, the method mayadditionally comprise: (c) growing a progeny plant of a subsequentgeneration from said seed of a progeny plant of a subsequent generationand crossing the progeny plant of a subsequent generation with itself ora second plant; and repeating the steps for an additional 2-10generations to produce a celery plant derived from the celery cultivarTBG 45. The plant derived from celery cultivar TBG 45 may be an inbredline, and the aforementioned repeated crossing steps may be defined ascomprising sufficient inbreeding to produce the inbred line. In themethod, it may be desirable to select particular plants resulting fromstep (c) for continued crossing according to steps (b) and (c). Byselecting plants having one or more desirable traits, a plant derivedfrom celery cultivar TBG 45 is obtained which possesses some of thedesirable traits of the line as well as potentially other selectedtraits. Also provided by the invention is a plant produced by this andthe other methods of the invention.

In another embodiment of the invention, the method of producing a celeryplant derived from the celery cultivar TBG 45 further comprises: (a)crossing the celery cultivar TBG 45-derived celery plant with itself oranother celery plant to yield additional celery cultivar TBG 45-derivedprogeny celery seed; (b) growing the progeny celery seed of step (a)under plant growth conditions to yield additional celery cultivar TBG45-derived celery plants; and (c) repeating the crossing and growingsteps of (a) and (b) to generate further celery cultivar TBG 45-derivedcelery plants. In specific embodiments, steps (a) and (b) may berepeated at least 1, 2, 3, 4, or 5 or more times as desired. Theinvention still further provides a celery plant produced by this and theforegoing methods.

Progeny of celery cultivar TBG 45 may also be characterized throughtheir filial relationship with celery cultivar TBG 45, as for example,being within a certain number of breeding crosses of celery cultivar TBG45. A breeding cross is a cross made to introduce new genetics into theprogeny, and is distinguished from a cross, such as a self or a sibcross, made to select among existing genetic alleles. The lower thenumber of breeding crosses in the pedigree, the closer the relationshipbetween celery cultivar TBG 45 and its progeny. For example, progenyproduced by the methods described herein may be within 1, 2, 3, 4 or 5breeding crosses of celery cultivar TBG 45.

One of ordinary skill in the art of plant breeding would know how toevaluate the traits of two plant varieties to determine if there is nosignificant difference between the two traits expressed by thosevarieties. For example, see, Fehr and Walt, Principles of CultivarDevelopment, pp. 261-286 (1987). Thus the invention includes celerycultivar TBG 45 progeny celery plants comprising a combination of atleast two cultivar TBG 45 traits selected from the group consisting ofthose listed in Table 1 or the cultivar TBG 45 combination of traitslisted in the Detailed Description of the Invention, so that saidprogeny celery plant is not significantly different for said traits thancelery cultivar TBG 45 as determined at the 5% significance level whengrown in the same environmental conditions. Using techniques describedherein, molecular markers may be used to identify said progeny plant asa celery cultivar TBG 45 progeny plant. Mean trait values may be used todetermine whether trait differences are significant, and preferably thetraits are measured on plants grown under the same environmentalconditions. Once such a variety is developed its value is substantialsince it is important to advance the germplasm base as a whole in orderto maintain or improve traits such as yield, disease resistance, pestresistance, and plant performance in extreme environmental conditions.

The goal of celery plant breeding is to develop new, unique, andsuperior celery cultivars. The breeder initially selects and crosses twoor more parental lines, followed by repeated selfing and selection,producing many new genetic combinations. The breeder can theoreticallygenerate billions of different genetic combinations via crossing,selfing, and mutations. The breeder has no direct control at thecellular level and the cultivars that are developed are unpredictable.This unpredictability is because the breeder's selection occurs inunique environments, with no control at the DNA level (usingconventional breeding procedures), and with millions of differentpossible genetic combinations being generated. A breeder of ordinaryskill in the art cannot predict the final resulting lines he develops,except possibly in a very gross and general fashion. The same breedercannot produce the same line twice by using the exact same originalparents and the same selection techniques. Therefore, two breeders willnever develop the same line, or even very similar lines, having the samecelery traits.

Pedigree breeding is used commonly for the improvement ofself-pollinating crops or inbred lines of cross-pollinating crops.Pedigree breeding starts with the crossing of two genotypes, such ascelery cultivar TBG 45 or a celery variety having all of themorphological and physiological characteristics of TBG 45, and anothercelery variety having one or more desirable characteristics that islacking or which complements celery cultivar TBG 45. If the two originalparents do not provide all the desired characteristics, other sourcescan be included in the breeding population. In the pedigree method,superior plants are selfed and selected in successive filialgenerations. In the succeeding filial generations, the heterozygouscondition gives way to the homozygous allele condition as a result ofinbreeding. Typically in the pedigree method of breeding, five or moresuccessive filial generations of selfing and selection is practiced: F₁to F₂; F₂ to F₃; F₃ to F₄; F₄ to F₅; etc. In some examples, 1, 2, 3, 4,5, 6, 7, 8, 9, 10, or more generations of selfing and selection arepracticed. After a sufficient amount of inbreeding, successive filialgenerations will serve to increase seed of the developed variety.Preferably, the developed variety comprises homozygous alleles at about95% or more of its loci.

In addition to being used to create backcross conversion populations,backcrossing can also be used in combination with pedigree breeding. Asdiscussed previously, backcrossing can be used to transfer one or morespecifically desirable traits from one variety (the donor parent) to adeveloped variety (the recurrent parent), which has good overallagronomic characteristics yet may lack one or more other desirabletraits. However, the same procedure can be used to move the progenytoward the genotype of the recurrent parent but at the same time retainmany components of the non-recurrent parent by stopping the backcrossingat an early stage and proceeding with selfing and selection. Forexample, a celery variety may be crossed with another variety to producea first generation progeny plant. The first generation progeny plant maythen be backcrossed to one of its parent varieties to create a F₁BC₁.Progeny are selfed and selected so that the newly developed variety hasmany of the attributes of the recurrent parent and yet several of thedesired attributes of the donor parent. This approach leverages thevalue and strengths of both parents for use in new celery varieties.

Therefore, in some examples a method of making a backcross conversion ofcelery cultivar TBG 45, comprising the steps of crossing a plant ofcelery cultivar TBG 45 or a celery variety having all of themorphological and physiological characteristics of TBG 45 with a donorplant possessing a desired trait to introduce the desired trait,selecting an F₁ progeny plant containing the desired trait, andbackcrossing the selected F₁ progeny plant to a plant of celery cultivarTBG 45 are provided. This method may further comprise the step ofobtaining a molecular marker profile of celery cultivar TBG 45 and usingthe molecular marker profile to select for a progeny plant with thedesired trait and the molecular marker profile of TBG 45. The molecularmarker profile can comprise information from one or more markers. In oneexample the desired trait is a mutant gene or transgene present in thedonor parent. In another example, the desired trait is a native trait inthe donor parent.

Mass and recurrent selections can be used to improve populations ofeither self- or cross-pollinating crops. A genetically variablepopulation of heterozygous individuals is either identified or createdby intercrossing several different parents. The best plants are selectedbased on individual superiority, outstanding progeny, or excellentcombining ability. The selected plants are intercrossed to produce a newpopulation in which further cycles of selection are continued.

The single-seed descent procedure in the strict sense refers to plantinga segregating population, harvesting a sample of one seed per plant, andusing the one-seed sample to plant the next generation. When thepopulation has been advanced from the F₂ to the desired level ofinbreeding, the plants from which lines are derived will each trace todifferent F₂ individuals. The number of plants in a population declineseach generation due to failure of some seeds to germinate or some plantsto produce at least one seed. As a result, not all of the F₂ plantsoriginally sampled in the population, will be represented by a progenywhen generation advance is completed.

Mutation breeding is another method of introducing new traits intocelery varieties. Mutations that occur spontaneously or are artificiallyinduced can be useful sources of variability for a plant breeder. Thegoal of artificial mutagenesis is to increase the rate of mutation for adesired characteristic. Mutation rates can be increased by manydifferent means including temperature, long-term seed storage, tissueculture conditions, radiation (such as X-rays, Gamma rays, neutrons,Beta radiation, or ultraviolet radiation), chemical mutagens (such asbase analogs like 5-bromo-uracil), antibiotics, alkylating agents (suchas sulfur mustards, nitrogen mustards, epoxides, ethyleneamines,sulfates, sulfonates, sulfones, or lactones), azide, hydroxylamine,nitrous acid or acridines. Once a desired trait is observed throughmutagenesis the trait may then be incorporated into existing germplasmby traditional breeding techniques. Details of mutation breeding can befound in “Principles of Cultivar Development” by Fehr, MacmillanPublishing Company, 1993. In addition, mutations created in other celeryplants may be used to produce a backcross conversion of celery cultivarTBG 45 that comprises such mutation.

Selection of celery plants for breeding is not necessarily dependent onthe phenotype of a plant and instead can be based on geneticinvestigations. For example, one may utilize a suitable genetic markerwhich is closely associated with a trait of interest. One of thesemarkers may therefore be used to identify the presence or absence of atrait in the offspring of a particular cross, and hence may be used inselection of progeny for continued breeding. This technique may commonlybe referred to as marker assisted selection. Any other type of geneticmarker or other assay which is able to identify the relative presence orabsence of a trait of interest in a plant may also be useful forbreeding purposes. Procedures for marker assisted selection applicableto the breeding of celeryes are well known in the art. Such methods willbe of particular utility in the case of recessive traits and variablephenotypes, or where conventional assays may be more expensive, timeconsuming or otherwise disadvantageous. Types of genetic markers whichcould be used in accordance with the invention include, but are notnecessarily limited to, Isozyme Electrophoresis, Restriction FragmentLength Polymorphisms (RFLPs), Simple Sequence Length Polymorphisms(SSLPs) (Williams et al., Nucleic Acids Res., 18:6531-6535, 1990),Randomly Amplified Polymorphic DNAs (RAPDs), DNA AmplificationFingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs),Arbitrary Primed Polymerase Chain Reaction (AP-PCR), Amplified FragmentLength Polymorphisms (AFLPs) (EP 534 858, specifically incorporatedherein by reference in its entirety), Simple Sequence Repeats (SSRs),and Single Nucleotide Polymorphisms (SNPs) (Wang et al., Science,280:1077-1082, 1998).

The production of double haploids can also be used for the developmentof homozygous varieties in a breeding program. Double haploids areproduced by the doubling of a set of chromosomes from a heterozygousplant to produce a completely homozygous individual. For example, see,Wan, et al., “Efficient Production of Doubled Haploid Plants ThroughColchicine Treatment of Anther-Derived Maize Callus,” Theoretical andApplied Genetics, 77:889-892 (1989) and U.S. Pat. No. 7,135,615. Thiscan be advantageous because the process omits the generations of selfingneeded to obtain a homozygous plant from a heterozygous source.

Descriptions of other breeding methods that are commonly used fordifferent traits and crops can be found in one of several referencebooks (e.g., Allard, “Principles of plant breeding,” John Wiley & Sons,NY, University of California, Davis, Calif, 50-98, 1960; Simmonds,“Principles of crop improvement,” Longman, Inc., NY, 369-399, 1979;Sneep and Hendriksen, “Plant breeding perspectives,” Wageningen (ed),Center for Agricultural Publishing and Documentation, 1979; Fehr, In:Soybeans: Improvement, Production and Uses,” 2d Ed., Manograph 16:249,1987; Fehr, “Principles of cultivar development,” Theory and Technique(Vol 1) and Crop Species Soybean (Vol 2), Iowa State Univ., MacmillianPub. Co., NY, 360-376, 1987; Poehlman and Sleper, “Breeding Field Crops”Iowa State University Press, Ames, 1995; Sprague and Dudley, eds., Cornand Improvement, 5th ed., 2006).

Genotypic Profile of TBG 45 and Progeny

In addition to phenotypic observations, a plant can also be identifiedby its genotype. The genotype of a plant can be characterized through agenetic marker profile which can identify plants of the same variety ora related variety, or which can be used to determine or validate apedigree. Genetic marker profiles can be obtained by techniques such asrestriction fragment length polymorphisms (RFLPs), randomly amplifiedpolymorphic DNAs (RAPDs), arbitrarily primed polymerase chain reaction(AP-PCR), DNA amplification fingerprinting (DAF), sequence characterizedamplified regions (SCARs), amplified fragment length polymorphisms(AFLPs), simple sequence repeats (SSRs) also referred to asmicrosatellites, single nucleotide polymorphisms (SNPs), or genome-wideevaluations such as genotyping-by-sequencing (GBS). For example, seeCregan et al. (1999) “An Integrated Genetic Linkage Map of the SoybeanGenome” Crop Science 39:1464-1490, and Berry et al. (2003) “AssessingProbability of Ancestry Using Simple Sequence Repeat Profiles:Applications to Maize Inbred Lines and Soybean Varieties” Genetics165:331-342, each of which are incorporated by reference herein in theirentirety. Favorable genotypes and or marker profiles, optionallyassociated with a trait of interest, may be identified by one or moremethodologies.

In some examples one or more markers are used, including but not limitedto AFLPs, RFLPs, ASH, SSRs, SNPs, indels, padlock probes, molecularinversion probes, microarrays, sequencing, and the like. In somemethods, a target nucleic acid is amplified prior to hybridization witha probe. In other cases, the target nucleic acid is not amplified priorto hybridization, such as methods using molecular inversion probes (see,for example Hardenbol et al. (2003) Nat Biotech 21:673-678). In someexamples, the genotype related to a specific trait is monitored, whilein other examples, a genome-wide evaluation including but not limited toone or more of marker panels, library screens, association studies,microarrays, gene chips, expression studies, or sequencing such aswhole-genome resequencing and genotyping-by-sequencing (GBS) may beused. In some examples, no target-specific probe is needed, for exampleby using sequencing technologies, including but not limited tonext-generation sequencing methods (see, for example, Metzker (2010) NatRev Genet 11:31-46; and, Egan et al. (2012) Am J Bot 99:175-185) such assequencing by synthesis (e.g., Roche 454 pyrosequencing, Illumina GenomeAnalyzer, and Ion Torrent PGM or Proton systems), sequencing by ligation(e.g., SOLiD from Applied Biosystems, and Polnator system from AzcoBiotech), and single molecule sequencing (SMS or third-generationsequencing) which eliminate template amplification (e.g., Helicossystem, and PacBio RS system from Pacific BioSciences). Furthertechnologies include optical sequencing systems (e.g., Starlight fromLife Technologies), and nanopore sequencing (e.g., GridION from OxfordNanopore Technologies). Each of these may be coupled with one or moreenrichment strategies for organellar or nuclear genomes in order toreduce the complexity of the genome under investigation via PCR,hybridization, restriction enzyme (see, e.g., Elshire et al. (2011) PLoSONE 6:e19379), and expression methods. In some examples, no referencegenome sequence is needed in order to complete the analysis.

With any of the genotyping techniques mentioned herein, polymorphismsmay be detected when the genotype and/or sequence of the plant ofinterest is compared to the genotype and/or sequence of one or morereference plants. The polymorphism revealed by these techniques may beused to establish links between genotype and phenotype. Thepolymorphisms may thus be used to predict or identify certain phenotypiccharacteristics, individuals, or even species. The polymorphisms aregenerally called markers. It is common practice for the skilled artisanto apply molecular DNA techniques for generating polymorphisms andcreating markers. The polymorphisms of this invention may be provided ina variety of mediums to facilitate use, e.g. a database or computerreadable medium, which may also contain descriptive annotations in aform that allows a skilled artisan to examine or query the polymorphismsand obtain useful information.

The invention further provides a method of determining the genotype of aplant of celery cultivar TBG 45, or a first generation progeny thereof,comprising detecting in the genome of the plant at least a firstpolymorphism. The method may, in certain embodiments, comprise detectinga plurality of polymorphisms in the genome of the plant. The method mayfurther comprise storing the results of the step of detecting theplurality of polymorphisms on a computer readable medium. The pluralityof polymorphisms are indicative of and/or give rise to the expression ofthe morphological and physiological characteristics of celery cultivarTBG 45. The invention further provides a computer readable mediumproduced by such a method.

In some examples, a plant, a plant part, or a seed of celery cultivarTBG 45 may be characterized by producing a molecular profile. Amolecular profile may include, but is not limited to, one or moregenotypic and/or phenotypic profile(s). A genotypic profile may include,but is not limited to, a marker profile, such as a genetic map, alinkage map, a trait maker profile, a SNP profile, an SSR profile, agenome-wide marker profile, a haplotype, and the like. A molecularprofile may also be a nucleic acid sequence profile, and/or a physicalmap. A phenotypic profile may include, but is not limited to, a proteinexpression profile, a metabolic profile, an mRNA expression profile, andthe like.

SSR technology is currently the most efficient and practical markertechnology; more marker loci can be routinely used and more alleles permarker locus can be found using SSRs in comparison to RFLPs. Forexample, Diwan and Cregan described a highly polymorphic microsatellitelocus in soybean with as many as 26 alleles. Diwan, N. and Cregan, P.B., Theor. Appl. Genet., 95:22-225 (1997). SNPs may also be used toidentify the unique genetic composition of the invention and progenyvarieties retaining that unique genetic composition. Various molecularmarker techniques may be used in combination to enhance overallresolution.

Molecular markers, which include markers identified through the use oftechniques such as Isozyme Electrophoresis, RFLPs, RAPDs, AP-PCR, DAF,SCARs, AFLPs, SSRs, and SNPs, may be used in plant breeding. One use ofmolecular markers is Quantitative Trait Loci (QTL) mapping. QTL mappingis the use of markers which are known to be closely linked to allelesthat have measurable effects on a quantitative trait. Selection in thebreeding process is based upon the accumulation of markers linked to thepositive effecting alleles and/or the elimination of the markers linkedto the negative effecting alleles from the plant's genome.

Molecular markers can also be used during the breeding process for theselection of qualitative traits. For example, markers closely linked toalleles or markers containing sequences within the actual alleles ofinterest can be used to select plants that contain the alleles ofinterest during a backcrossing breeding program. The markers can also beused to select toward the genome of the recurrent parent and against themarkers of the donor parent. This procedure attempts to minimize theamount of genome from the donor parent that remains in the selectedplants. It can also be used to reduce the number of crosses back to therecurrent parent needed in a backcrossing program. The use of molecularmarkers in the selection process is often called genetic marker enhancedselection or marker-assisted selection. Molecular markers may also beused to identify and exclude certain sources of germplasm as parentalvarieties or ancestors of a plant by providing a means of trackinggenetic profiles through crosses.

Particular markers used for these purposes are not limited to the set ofmarkers disclosed herein, but may include any type of marker and markerprofile which provides a means of distinguishing varieties. In additionto being used for identification of celery cultivar TBG 45, a hybridproduced through the use of TBG 45, and the identification orverification of pedigree for progeny plants produced through the use ofTBG 45, a genetic marker profile is also useful in developing a geneconversion of TBG 45.

Means of performing genetic marker profiles using SNP and SSRpolymorphisms are well known in the art. SNPs are genetic markers basedon a polymorphism in a single nucleotide. A marker system based on SNPscan be highly informative in linkage analysis relative to other markersystems in that multiple alleles may be present.

The SSR profile of celery cultivar TBG 45 can be used to identify plantscomprising celery cultivar TBG 45 as a parent, since such plants willcomprise the same homozygous alleles as celery cultivar TBG 45. Becausethe celery variety is essentially homozygous at all relevant loci, mostloci should have only one type of allele present. In contrast, a geneticmarker profile of an F₁ progeny should be the sum of those parents,e.g., if one parent was homozygous for allele x at a particular locus,and the other parent homozygous for allele y at that locus, then the F₁progeny will be xy (heterozygous) at that locus. Subsequent generationsof progeny produced by selection and breeding are expected to be ofgenotype x (homozygous), y (homozygous), or xy (heterozygous) for thatlocus position. When the F₁ plant is selfed or sibbed for successivefilial generations, the locus should be either x or y for that position.

In addition, plants and plant parts substantially benefiting from theuse of celery cultivar TBG 45 in their development, such as celerycultivar TBG 45 comprising a gene conversion, backcross conversion,transgene, or genetic sterility factor, may be identified by having amolecular marker profile with a high percent identity to celery cultivarTBG 45. Such a percent identity might be 95%, 96%, 97%, 98%, 99%, 99.5%,or 99.9% identical to celery cultivar TBG 45.

The SSR profile of celery cultivar TBG 45 can also be used to identifyessentially derived varieties and other progeny varieties developed fromthe use of celery cultivar TBG 45, as well as cells and other plantparts thereof. Such plants may be developed using the markers identifiedin WO 00/31964, U.S. Pat. Nos. 6,162,967, and 7,288,386. Progeny plantsand plant parts produced using celery cultivar TBG 45 may be identifiedby having a molecular marker profile of at least 25%, 30%, 35%, 40%,45%, 50%, 55%, 60%, 65%, 70%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%,83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98%, 99%, or 99.5% genetic contribution from celery cultivar TBG45, as measured by either percent identity or percent similarity. Suchprogeny may be further characterized as being within a pedigree distanceof celery cultivar TBG 45, such as within 1, 2, 3, 4, or 5 or lesscross-pollinations to a celery plant other than celery cultivar TBG 45or a plant that has celery cultivar TBG 45 as a progenitor. Uniquemolecular profiles may be identified with other molecular tools such asSNPs and RFLPs.

While determining the genotypic profile of the plants described supra,several unique SSR profiles may also be identified which did not appearin either parent of such plant. Such unique SSR profiles may ariseduring the breeding process from recombination or mutation. Acombination of several unique alleles provides a means of identifying aplant variety, an F₁ progeny produced from such variety, and progenyproduced from such variety.

Molecular data from TBG 45 may be used in a plant breeding process.Nucleic acids may be isolated from a seed of TBG 45 or from a plant,plant part, or cell produced by growing a seed of TBG 45, or from a seedof TBG 45 with a gene conversion, or from a plant, plant part, or cellof TBG 45 with a gene conversion. One or more polymorphisms may beisolated from the nucleic acids. A plant having one or more of theidentified polymorphisms may be selected and used in a plant breedingmethod to produce another plant.

In another embodiment of the invention, the genetic complement of thecelery cultivar TBG 45 is provided. The phrase “genetic complement” isused to refer to the aggregate of nucleotide sequences, the expressionof which sequences defines the phenotype of, in the present case, acelery plant, or a cell or tissue of that plant. A genetic complementthus represents the genetic makeup of a cell, tissue or plant, and ahybrid genetic complement represents the genetic makeup of a hybridcell, tissue or plant. The invention thus provides celery plant cellsthat have a genetic complement in accordance with the celery plant cellsdisclosed herein, and plants, seeds and plants containing such cells.Plant genetic complements may be assessed by genetic marker profiles,and by the expression of phenotypic traits that are characteristic ofthe expression of the genetic complement, e.g., isozyme typing profiles.

Introduction of a New Trait or Locus into Celery Cultivar TBG 45

Cultivar TBG 45 represents a new base genetic variety into which a newgene, locus or trait may be introgressed. Backcrossing and directtransformation represent two important methods that can be used toaccomplish such an introgression.

Single Gene (Locus) Conversions

When the term “celery plant” is used in the context of the presentinvention, this also includes any single gene or locus conversions ofthat variety. The term “single locus converted plant” or “single geneconverted plant” refers to those celery plants which are developed bybackcrossing or genetic engineering, wherein essentially all of thedesired morphological and physiological characteristics of a variety arerecovered in addition to the one or more genes transferred into thevariety via the backcrossing technique or genetic engineering.Backcrossing methods can be used with the present invention to improveor introduce a characteristic into the variety.

A backcross conversion of celery cultivar TBG 45 occurs when DNAsequences are introduced through backcrossing (Hallauer, et al., “CornBreeding,” Corn and Corn Improvements, No. 18, pp. 463-481 (1988)), withcelery cultivar TBG 45 utilized as the recurrent parent. Both naturallyoccurring and transgenic DNA sequences may be introduced throughbackcrossing techniques. A backcross conversion may produce a plant witha trait or locus conversion in at least two or more backcrosses,including at least 2 crosses, at least 3 crosses, at least 4 crosses, atleast 5 crosses, and the like. Molecular marker assisted breeding orselection may be utilized to reduce the number of backcrosses necessaryto achieve the backcross conversion. For example, see, Openshaw, S. J.,et al., Marker-assisted Selection in Backcross Breeding, ProceedingsSymposium of the Analysis of Molecular Data, Crop Science Society ofAmerica, Corvallis, Oregon (August 1994), where it is demonstrated thata backcross conversion can be made in as few as two backcrosses.

The complexity of the backcross conversion method depends on the type oftrait being transferred (single genes or closely linked genes ascompared to unlinked genes), the level of expression of the trait, thetype of inheritance (cytoplasmic or nuclear), and the types of parentsincluded in the cross. It is understood by those of ordinary skill inthe art that for single gene traits that are relatively easy toclassify, the backcross method is effective and relatively easy tomanage. (See, Hallauer, et al., Corn and Corn Improvement, Sprague andDudley, Third Ed. (1998)). Desired traits that may be transferredthrough backcross conversion include, but are not limited to, sterility(nuclear and cytoplasmic), fertility restoration, nutritionalenhancements, drought tolerance, nitrogen utilization, altered fattyacid profile, modified fatty acid metabolism, modified carbohydratemetabolism, industrial enhancements, yield stability, yield enhancement,disease resistance (bacterial, fungal, or viral), insect resistance, andherbicide resistance. In addition, an introgression site itself, such asan FRT site, Lox site, or other site specific integration site, may beinserted by backcrossing and utilized for direct insertion of one ormore genes of interest into a specific plant variety.

A single locus may contain several transgenes, such as a transgene fordisease resistance that, in the same expression vector, also contains atransgene for herbicide resistance. The gene for herbicide resistancemay be used as a selectable marker and/or as a phenotypic trait. Asingle locus conversion of site specific integration system allows forthe integration of multiple genes at a known recombination site in thegenome. At least one, at least two or at least three and less than ten,less than nine, less than eight, less than seven, less than six, lessthan five or less than four locus conversions may be introduced into theplant by backcrossing, introgression or transformation to express thedesired trait, while the plant, or a plant grown from the seed, plantpart or plant cell, otherwise retains the phenotypic characteristics ofthe deposited seed when grown under the same environmental conditions.

The backcross conversion may result from either the transfer of adominant allele or a recessive allele. Selection of progeny containingthe trait of interest is accomplished by direct selection for a traitassociated with a dominant allele. Transgenes transferred viabackcrossing typically function as a dominant single gene trait and arerelatively easy to classify. Selection of progeny for a trait that istransferred via a recessive allele requires growing and selfing thefirst backcross generation to determine which plants carry the recessivealleles. Recessive traits may require additional progeny testing insuccessive backcross generations to determine the presence of the locusof interest. The last backcross generation is usually selfed to givepure breeding progeny for the gene(s) being transferred, although abackcross conversion with a stably introgressed trait may also bemaintained by further backcrossing to the recurrent parent withselection for the converted trait.

Along with selection for the trait of interest, progeny are selected forthe phenotype of the recurrent parent. The backcross is a form ofinbreeding, and the features of the recurrent parent are automaticallyrecovered after successive backcrosses. Poehlman, Breeding Field Crops,p. 204 (1987). Poehlman suggests from one to four or more backcrosses,but as noted above, the number of backcrosses necessary can be reducedwith the use of molecular markers. Other factors, such as a geneticallysimilar donor parent, may also reduce the number of backcrossesnecessary. As noted by Poehlman, backcrossing is easiest for simplyinherited, dominant, and easily recognized traits.

One process for adding or modifying a trait or locus in celery cultivarTBG 45 comprises crossing celery cultivar TBG 45 plants grown fromcelery cultivar TBG 45 seed with plants of another celery variety thatcomprise the desired trait, gene or locus, selecting F₁ progeny plantsthat comprise the desired trait, gene or locus to produce selected F₁progeny plants, crossing the selected progeny plants with the celerycultivar TBG 45 plants to produce backcross progeny plants, selectingfor backcross progeny plants that have the desired trait, gene or locusand the morphological characteristics of celery cultivar TBG 45 toproduce selected backcross progeny plants, and backcrossing to celerycultivar TBG 45 three or more times in succession to produce selectedfourth or higher backcross progeny plants that comprise said trait, geneor locus. The modified celery cultivar TBG 45 may be furthercharacterized as having the physiological and morphologicalcharacteristics of celery cultivar TBG 45 as determined at the 5%significance level when grown in the same environmental conditionsand/or may be characterized by percent similarity or identity to celerycultivar TBG 45 as determined by SSR markers. The above method may beutilized with fewer backcrosses in appropriate situations, such as whenthe donor parent is highly related or markers are used in the selectionstep. Desired traits that may be used include those nucleic acids knownin the art, some of which are listed herein, that will affect traitsthrough nucleic acid expression or inhibition. Desired loci include theintrogression of FRT, Lox, and other sites for site specificintegration, which may also affect a desired trait if a functionalnucleic acid is inserted at the integration site.

In addition, the above process and other similar processes describedherein may be used to produce first generation progeny celery seed byadding a step at the end of the process that comprises crossing celerycultivar TBG 45 with the introgressed trait or locus with a differentcelery plant and harvesting the resultant first generation progenycelery seed.

Methods for Genetic Engineering of Celery

With the advent of molecular biological techniques that have allowed theisolation and characterization of genes that encode specific proteinproducts, scientists in the field of plant biology developed a stronginterest in engineering the genome of plants (genetic engineering) tocontain and express foreign genes, or additional, or modified versionsof native, or endogenous genes (perhaps driven by different promoters)in order to alter the traits of a plant in a specific manner. Plantsaltered by genetic engineering are often referred to as ‘geneticallymodified’. Any DNA sequences, whether from a different species or fromthe same species, which are introduced into the genome usingtransformation and/or various breeding methods, are referred to hereincollectively as “transgenes.” Over the last fifteen to twenty yearsseveral methods for producing transgenic plants have been developed, andthe present invention, in particular embodiments, also relates totransformed versions of the claimed cultivar.

Vectors used for the transformation of celery cells are not limited solong as the vector can express an inserted DNA in the cells. Forexample, vectors comprising promoters for constitutive gene expressionin celery cells (e.g., cauliflower mosaic virus 35S promoter) andpromoters inducible by exogenous stimuli can be used. Examples ofsuitable vectors include pBI binary vector. The “celery cell” into whichthe vector is to be introduced includes various forms of celery cells,such as cultured cell suspensions, protoplasts, leaf sections, andcallus. A vector can be introduced into celery cells by known methods,such as the polyethylene glycol method, polycation method,electroporation, Agrobacterium-mediated transfer, particle bombardmentand direct DNA uptake by protoplasts. See, e.g., Pang et al. (The PlantJ., 9, 899-909, 1996).

Numerous methods for plant transformation have been developed, includingbiological and physical, plant transformation protocols. See, forexample, Miki, et al., “Procedures for Introducing Foreign DNA intoPlants” in Methods in Plant Molecular Biology and Biotechnology, Glickand Thompson (Eds.), CRC Press, Inc., Boca Raton, pp. 67-88 (1993). Inaddition, expression vectors and in vitro culture methods for plant cellor tissue transformation and regeneration of plants are available. See,for example, Gruber, et al., “Vectors for Plant Transformation” inMethods in Plant Molecular Biology and Biotechnology, Glick and Thompson(Eds.), CRC Press, Inc., Boca Raton, pp. 89-119 (1993).

A. Agrobacterium-Mediated Transformation:

One method for introducing an expression vector into plants is based onthe natural transformation system of Agrobacterium. See, for example,Horsch, et al., Science, 227:1229 (1985). A. tumefaciens and A.rhizogenes are plant pathogenic soil bacteria which geneticallytransform plant cells. The Ti and Ri plasmids of A. tumefaciens and A.rhizogenes, respectively, carry genes responsible for genetictransformation of the plant. See, for example, Kado, C. I., Crit. Rev.Plant Sci., 10:1 (1991). Descriptions of Agrobacterium vector systemsand methods for Agrobacterium-mediated gene transfer are provided byGruber, et al., supra, Miki, et al., supra, and Moloney, et al., PlantCell Rep., 8:238 (1989). See also, U.S. Pat. No. 5,563,055 (Townsend andThomas), issued Oct. 8, 1996.

Agrobacterium-mediated transfer is a widely applicable system forintroducing gene loci into plant cells, including celery. An advantageof the technique is that DNA can be introduced into whole plant tissues,thereby bypassing the need for regeneration of an intact plant from aprotoplast. Modern Agrobacterium transformation vectors are capable ofreplication in E. coli as well as Agrobacterium, allowing for convenientmanipulations (Klee et al., Bio. Tech., 3(7):637-642, 1985). Moreover,recent technological advances in vectors for Agrobacterium-mediated genetransfer have improved the arrangement of genes and restriction sites inthe vectors to facilitate the construction of vectors capable ofexpressing various polypeptide coding genes. The vectors described haveconvenient multi-linker regions flanked by a promoter and apolyadenylation site for direct expression of inserted polypeptidecoding genes. Additionally, Agrobacterium containing both armed anddisarmed Ti genes can be used for transformation.

In those plant strains where Agrobacterium-mediated transformation isefficient, it is the method of choice because of the facile and definednature of the gene locus transfer. The use of Agrobacterium-mediatedplant integrating vectors to introduce DNA into plant cells is wellknown in the art (Fraley et al., Bio. Tech., 3(7):629-635, 1985; U.S.Pat. No. 5,563,055). For example, U.S. Pat. No. 5,349,124 describes amethod of transforming celery plant cells using Agrobacterium-mediatedtransformation. By inserting a chimeric gene having a DNA codingsequence encoding for the full-length B.t. toxin protein that expressesa protein toxic toward Lepidopteran larvae, this methodology resulted incelery having resistance to such insects.

B. Direct Gene Transfer:

Several methods of plant transformation, collectively referred to asdirect gene transfer, have been developed as an alternative toAgrobacterium-mediated transformation. A generally applicable method fordelivering transforming DNA segments to plant cells ismicroprojectile-mediated transformation, or microprojectile bombardment.In this method, particles are coated with nucleic acids and deliveredinto cells by a propelling force. Sanford, et al., Part. Sci. Technol.,5:27 (1987); Sanford, J. C., Trends Biotech., 6:299 (1988); Klein, etal., Bio/technology, 6:559-563 (1988); Sanford, J. C., Physiol Plant,7:206 (1990); Klein, et al., Bio/technology, 10:268 (1992). See also,U.S. Pat. No. 5,015,580 (Christou, et al.), issued May 14, 1991; U.S.Pat. No. 5,322,783 (Tomes, et al.), issued Jun. 21, 1994.

Another method for physical delivery of DNA to plants is sonication oftarget cells. Zhang, et al., Bio/technology, 9:996 (1991).Alternatively, liposome and spheroplast fusion have been used tointroduce expression vectors into plants. Deshayes, et al., EMBO 1,4:2731 (1985); Christou, et al., PNAS, 84:3962 (1987). Direct uptake ofDNA into protoplasts using CaCl₂) precipitation, calcium phosphateprecipitation, polyethylene glycol treatment, polyvinyl alcohol, orpoly-L-ornithine has also been reported. See, e.g., Potrykus et al.,Mol. Gen. Genet., 199:183-188, 1985; Omirulleh et al., Plant Mol. Biol21(3):415-428, 1993; Fromm et al., Nature, 312:791-793, 1986; Uchimiyaet al., Mol. Gen. Genet., 204:204, 1986; Marcotte et al., Nature,335:454, 1988; Hain, et al., Mol. Gen. Genet., 199:161, 1985 and Draper,et al., Plant Cell Physiol. 23:451, 1982.

Electroporation of protoplasts and whole cells and tissues has also beendescribed. Donn, et al., In Abstracts of VIIth International Congress onPlant Cell and Tissue Culture IAPTC, A2-38, p. 53, 1990; D'Halluin, etal., Plant Cell, 4:1495-1505, 1992; and Spencer, et al., Plant Mol.Biol., 24:51-61, 1994. Another illustrative embodiment of a method fordelivering DNA into plant cells by acceleration is the BiolisticsParticle Delivery System, which can be used to propel particles coatedwith DNA or cells through a screen, such as a stainless steel or Nytexscreen, onto a surface covered with target celery cells.

Transformation of plants and expression of foreign genetic elements isexemplified in Choi et al., Plant Cell Rep., 13: 344-348, 1994 and Ellulet al., Theor. Appl. Genet., 107:462-469, 2003.

Following transformation of celery target tissues, expression ofselectable marker genes allows for preferential selection of transformedcells, tissues, and/or plants, using regeneration and selection methodsnow well known in the art.

The methods described herein for transformation would typically be usedfor producing a transgenic variety. The transgenic variety could then becrossed, with another (non-transformed or transformed) variety, in orderto produce a new transgenic variety. Alternatively, a genetic traitwhich has been engineered into a particular celery cultivar using thetransformation techniques described could be moved into another cultivarusing traditional backcrossing techniques that are well known in theplant breeding arts. For example, a backcrossing approach could be usedto move an engineered trait from a public, non-elite variety into anelite variety, or from a variety containing a foreign gene in its genomeinto a variety or varieties which do not contain that gene. As usedherein, “crossing” can refer to a simple X by Y cross, or the process ofbackcrossing, depending on the context.

Expression Vectors for Celery Transformation: Marker Genes

Expression vectors include at least one genetic marker, operably linkedto a regulatory element (a promoter, for example) that allowstransformed cells containing the marker to be either recovered bynegative selection, i.e., inhibiting growth of cells that do not containthe selectable marker gene, or by positive selection, i.e., screeningfor the product encoded by the genetic marker. Many commonly usedselectable marker genes for plant transformation are well known in thetransformation arts, and include, for example, genes that code forenzymes that metabolically detoxify a selective chemical agent which maybe an antibiotic or a herbicide, or genes that encode an altered targetwhich is insensitive to the inhibitor. A few positive selection methodsare also known in the art.

One commonly used selectable marker gene for plant transformation is theneomycin phosphotransferase II (nptII) gene, isolated from transposonTn5, which when placed under the control of plant regulatory signalswhich confers resistance to kanamycin. Fraley et al., Proc. Natl. Acad.Sci. U.S.A., 80:4803 (1983). Another commonly used selectable markergene is the hygromycin phosphotransferase gene which confers resistanceto the antibiotic hygromycin. Vanden Elzen et al., Plant Mol. Biol.,5:299 (1985).

Additional selectable marker genes of bacterial origin that conferresistance to antibiotics include gentamycin acetyl transferase,streptomycin phosphotransferase, aminoglycoside-3′-adenyl transferase,the bleomycin resistance determinant. Hayford et al., Plant Physiol.86:1216 (1988), Jones et al., Mol. Gen. Genet., 210:86 (1987), Svab etal., Plant Mol. Biol. 14:197 (1990) Hille et al., Plant Mol. Biol. 7:171(1986). Other selectable marker genes confer resistance to herbicidessuch as glyphosate, glufosinate or bromoxynil. Comai et al., Nature317:741-744 (1985), Gordon-Kamm et al., Plant Cell 2:603-618 (1990) andStalker et al., Science 242:419-423 (1988). Selectable marker genes forplant transformation not of bacterial origin include, for example, mousedihydrofolate reductase, plant 5-enolpyruvylshikimate-3-phosphatesynthase and plant acetolactate synthase. Eichholtz et al., Somatic CellMol. Genet. 13:67 (1987), Shah et al., Science 233:478 (1986), Charestet al., Plant Cell Rep. 8:643 (1990).

Another class of marker genes for plant transformation requiresscreening of presumptively transformed plant cells rather than directgenetic selection of transformed cells for resistance to a toxicsubstance such as an antibiotic. These genes are particularly useful toquantify or visualize the spatial pattern of expression of a gene inspecific tissues and are frequently referred to as reporter genesbecause they can be fused to a gene or gene regulatory sequence for theinvestigation of gene expression. Commonly used genes for screeningpresumptively transformed cells include α-glucuronidase (GUS,α-galactosidase, luciferase and chloramphenicol, acetyltransferase.Jefferson, R. A., Plant Mol. Bio. Rep. 5:387 (1987), Teeri et al., EMBOJ. 8:343 (1989), Koncz et al., Proc. Natl. Acad. Sci U.S.A. 84:131(1987), DeBlock et al., EMBO J. 3:1681 (1984).

In vivo methods for visualizing GUS activity that do not requiredestruction of plant tissue are available. Molecular Probes publication2908, IMAGENE GREEN, p. 1-4 (1993) and Naleway et al., J. Cell Biol.115:151a (1991). However, these in vivo methods for visualizing GUSactivity have not proven useful for recovery of transformed cellsbecause of low sensitivity, high fluorescent backgrounds and limitationsassociated with the use of luciferase genes as selectable markers.

More recently, a gene encoding Green Fluorescent Protein (GFP) has beenutilized as a marker for gene expression in prokaryotic and eukaryoticcells. Chalfie et al., Science 263:802 (1994). GFP and mutants of GFPmay be used as screenable markers.

Expression Vectors for Celery Transformation: Promoters

Genes included in expression vectors must be driven by a nucleotidesequence comprising a regulatory element, for example, a promoter.Several types of promoters are now well known in the transformationarts, as are other regulatory elements that can be used alone or incombination with promoters.

As used herein, “promoter” includes reference to a region of DNAupstream from the start of transcription and involved in recognition andbinding of RNA polymerase and other proteins to initiate transcription.A “plant promoter” is a promoter capable of initiating transcription inplant cells. Examples of promoters under developmental control includepromoters that preferentially initiate transcription in certain tissues,such as leaves, roots, seeds, fibers, xylem vessels, tracheids, orsclerenchyma. Such promoters are referred to as “tissue-preferred”.Promoters which initiate transcription only in certain tissue arereferred to as “tissue-specific”. A “cell type” specific promoterprimarily drives expression in certain cell types in one or more organs,for example, vascular cells in roots or leaves. An “inducible” promoteris a promoter which is under environmental control. Examples ofenvironmental conditions that may effect transcription by induciblepromoters include anaerobic conditions or the presence of light.Tissue-specific, tissue-preferred, cell type specific, and induciblepromoters constitute the class of “non-constitutive” promoters. A“constitutive” promoter is a promoter which is active under mostenvironmental conditions.

A. Inducible Promoters:

An inducible promoter is operably linked to a gene for expression incelery. Optionally, the inducible promoter is operably linked to anucleotide sequence encoding a signal sequence which is operably linkedto a gene for expression in celery. With an inducible promoter the rateof transcription increases in response to an inducing agent.

Any inducible promoter can be used in the instant invention. See Ward etal., Plant Mol. Biol. 22:361-366 (1993). Exemplary inducible promotersinclude, but are not limited to, that from the ACEI system whichresponds to copper (Meft et al., PNAS 90:4567-4571 (1993)); In2 genefrom maize which responds to benzenesulfonamide herbicide safeners(Hershey et al., Mol. Gen Genetics 227:229-237 (1991) and Gatz et al.,Mol. Gen. Genetics 243:32-38 (1994)) or Tet repressor from Tn10 (Gatz etal., Mol. Gen. Genetics 227:229-237 (1991). A particularly preferredinducible promoter is a promoter that responds to an inducing agent towhich plants do not normally respond. An exemplary inducible promoter isthe inducible promoter from a steroid hormone gene, the transcriptionalactivity of which is induced by a glucocorticosteroid hormone. Schena etal., Proc. Natl. Acad. Sci. U.S.A. 88:0421 (1991).

B. Constitutive Promoters:

A constitutive promoter is operably linked to a gene for expression incelery or the constitutive promoter is operably linked to a nucleotidesequence encoding a signal sequence which is operably linked to a genefor expression in celery.

Many different constitutive promoters can be utilized in the instantinvention. Exemplary constitutive promoters include, but are not limitedto, the promoters from plant viruses such as the 35S promoter from CaMV(Odell et al., Nature 313:810-812 (1985) and the promoters from suchgenes as rice actin (McElroy et al., Plant Cell 2:163-171 (1990));ubiquitin (Christensen et al., Plant Mol. Biol. 12:619-632 (1989) andChristensen et al., Plant Mol. Biol. 18:675-689 (1992)); pEMU (Last etal., Theor. Appl. Genet. 81:581-588 (1991)); MAS (Velten et al., EMBO J.3:2723-2730 (1984)) and maize H3 histone (Lepetit et al., Mol. Gen.Genetics 231:276-285 (1992) and Atanassova et al., Plant Journal 2 (3):291-300 (1992)). The ALS promoter, Xba1/Nco1 fragment 5′ to the Brassicanapus ALS3 structural gene (or a nucleotide sequence similarity to saidXba1/Nco1 fragment), represents a particularly useful constitutivepromoter. See PCT application WO 96/30530.

C. Tissue-Specific or Tissue-Preferred Promoters:

A tissue-specific promoter is operably linked to a gene for expressionin celery. Optionally, the tissue-specific promoter is operably linkedto a nucleotide sequence encoding a signal sequence which is operablylinked to a gene for expression in celery. Plants transformed with agene of interest operably linked to a tissue-specific promoter producethe protein product of the transgene exclusively, or preferentially, ina specific tissue.

Any tissue-specific or tissue-preferred promoter can be utilized in theinstant invention. Exemplary tissue-specific or tissue-preferredpromoters include, but are not limited to, a root-preferred promoter,such as that from the phaseolin gene (Murai et al., Science 23:476-482(1983) and Sengupta-Gopalan et al., Proc. Natl. Acad. Sci. U.S.A.82:3320-3324 (1985)); a leaf-specific and light-induced promoter such asthat from cab or rubisco (Simpson et al., EMBO J. 4(11):2723-2729 (1985)and Timko et al., Nature 318:579-582 (1985)); an anther-specificpromoter such as that from LAT52 (Twell et al., Mol. Gen. Genetics217:240-245 (1989)); a pollen-specific promoter such as that from Zm13(Guerrero et al., Mol. Gen. Genetics 244:161-168 (1993)) or amicrospore-preferred promoter such as that from apg (Twell et al., Sex.Plant Reprod. 6:217-224 (1993)).

Signal Sequences for Targeting Proteins to Subcellular Compartments

Transport of protein produced by transgenes to a subcellular compartmentsuch as the chloroplast, vacuole, peroxisome, glyoxysome, cell wall ormitochondrion or for secretion into the apoplast, is accomplished bymeans of operably linking the nucleotide sequence encoding a signalsequence to the 5′ and/or 3′ region of a gene encoding the protein ofinterest. Targeting sequences at the 5′ and/or 3′ end of the structuralgene may determine, during protein synthesis and processing, where theencoded protein is ultimately compartmentalized.

The presence of a signal sequence directs a polypeptide to either anintracellular organelle or subcellular compartment or for secretion tothe apoplast. Many signal sequences are known in the art. See, forexample Becker et al., Plant Mol. Biol. 20:49 (1992), Close, P. S.,Master's Thesis, Iowa State University (1993), Knox, C., et al.,“Structure and Organization of Two Divergent Alpha-Amylase Genes fromBarley”, Plant Mol. Biol. 9:3-17 (1987), Lerner et al., Plant Physiol.91:124-129 (1989), Fontes et al., Plant Cell 3:483-496 (1991), Matsuokaet al., Proc. Natl. Acad. Sci. 88:834 (1991), Gould et al., J. Cell.Biol. 108:1657 (1989), Creissen et al., Plant J. 2:129 (1991), Kalderon,et al., A short amino acid sequence able to specify nuclear location,Cell 39:499-509 (1984), Steifel, et al., Expression of a maize cell wallhydroxyproline-rich glycoprotein gene in early leaf and root vasculardifferentiation, Plant Cell 2:785-793 (1990).

Additional Methods for Genetic Engineering of Celery

In general, methods to transform, modify, edit or alter plant endogenousgenomic DNA include altering the plant native DNA sequence or apre-existing transgenic sequence including regulatory elements, codingand non-coding sequences. These methods can be used, for example, totarget nucleic acids to pre-engineered target recognition sequences inthe genome. Such pre-engineered target sequences may be introduced bygenome editing or modification. As an example, a genetically modifiedplant variety is generated using “custom” or engineered endonucleasessuch as meganucleases produced to modify plant genomes (see e.g., WO2009/114321; Gao et al. (2010) Plant Journal 1:176-187). Anothersite-directed engineering method is through the use of zinc fingerdomain recognition coupled with the restriction properties ofrestriction enzyme. See e.g., Urnov, et al., (2010) Nat Rev Genet.11(9):636-46; Shukla, et al., (2009) Nature 459 (7245):437-41. Atranscription activator-like (TAL) effector-DNA modifying enzyme (TALEor TALEN) is also used to engineer changes in plant genome. See e.g.,US20110145940, Cermak et al., (2011) Nucleic Acids Res. 39(12) and Bochet al., (2009), Science 326(5959): 1509-12. Site-specific modificationof plant genomes can also be performed using the bacterial type IICRISPR (clustered regularly interspaced short palindromic repeats)/Cas(CRISPR-associated) system, as well as similar CRISPR relatedtechnologies. See e.g., Belhaj et al., (2013), Plant Methods 9: 39; TheCas9/guide RNA-based system allows targeted cleavage of genomic DNAguided by a customizable small noncoding RNA in plants (see e.g., WO2015026883A1, incorporated herein by reference).

A genetic map can be generated that identifies the approximatechromosomal location of an integrated DNA molecule, for example viaconventional restriction fragment length polymorphisms (RFLP),polymerase chain reaction (PCR) analysis, simple sequence repeats (SSR),and single nucleotide polymorphisms (SNP). For exemplary methodologiesin this regard, see Glick and Thompson, Methods in Plant MolecularBiology and Biotechnology, pp. 269-284 (CRC Press, Boca Raton, 1993).

Wang et al. discuss “Large Scale Identification, Mapping and Genotypingof Single-Nucleotide Polymorphisms in the Human Genome”, Science (1998)280:1077-1082, and similar capabilities are increasingly available forthe celery genome. Map information concerning chromosomal location isuseful for proprietary protection of a subject transgenic plant. Ifunauthorized propagation is undertaken and crosses made with othergermplasm, the map of the integration region can be compared to similarmaps for suspect plants to determine if the latter have a commonparentage with the subject plant. Map comparisons could involvehybridizations, RFLP, PCR, SSR, sequencing or combinations thereof, allof which are conventional techniques. SNPs may also be used alone or incombination with other techniques.

Celery Cultivar TBG 45 Further Comprising a Transgene

Transgenes and transformation methods provide means to engineer thegenome of plants to contain and express heterologous genetic elements,including but not limited to foreign genetic elements, additional copiesof endogenous elements, and/or modified versions of native or endogenousgenetic elements, in order to alter at least one trait of a plant in aspecific manner. Any heterologous DNA sequence(s), whether from adifferent species or from the same species, which are inserted into thegenome using transformation, backcrossing, or other methods known to oneof skill in the art are referred to herein collectively as transgenes.The sequences are heterologous based on sequence source, location ofintegration, operably linked elements, or any combination thereof. Oneor more transgenes of interest can be introduced into celery cultivarTBG 45. Transgenic variants of celery cultivar TBG 45 plants, seeds,cells, and parts thereof or derived therefrom are provided. Transgenicvariants of TBG 45 comprise the physiological and morphologicalcharacteristics of celery cultivar TBG 45, such as determined at the 5%significance level when grown in the same environmental conditions,and/or may be characterized or identified by percent similarity oridentity to TBG 45 as determined by SSR or other molecular markers. Insome examples, transgenic variants of celery cultivar TBG 45 areproduced by introducing at least one transgene of interest into celerycultivar TBG 45 by transforming TBG 45 with a polynucleotide comprisingthe transgene of interest. In other examples, transgenic variants ofcelery cultivar TBG 45 are produced by introducing at least onetransgene by introgressing the transgene into celery cultivar TBG 45 bycrossing.

In one example, a process for modifying celery cultivar TBG 45 with theaddition of a desired trait, said process comprising transforming acelery plant of cultivar TBG 45 with a transgene that confers a desiredtrait is provided. Therefore, transgenic TBG 45 celery cells, plants,plant parts, and seeds produced from this process are provided. In someexamples one more desired traits may include traits such as sterility(nuclear and cytoplasmic), fertility restoration, nutritionalenhancements, drought tolerance, nitrogen utilization, altered fattyacid profile, modified fatty acid metabolism, modified carbohydratemetabolism, industrial enhancements, yield stability, yield enhancement,disease resistance (bacterial, fungal, or viral), insect resistance, andherbicide resistance. The specific gene may be any known in the art orlisted herein, including but not limited to a polynucleotide conferringresistance to an ALS-inhibitor herbicide, imidazolinone, sulfonylurea,protoporphyrinogen oxidase (PPO) inhibitors, hydroxyphenyl pyruvatedioxygenase (HPPD) inhibitors, glyphosate, glufosinate, triazine,2,4-dichlorophenoxyacetic acid (2,4-D), dicamba, broxynil, metribuzin,or benzonitrile herbicides; a polynucleotide encoding a Bacillusthuringiensis polypeptide, a polynucleotide encoding a phytase, a fattyacid desaturase (e.g., FAD-2, FAD-3), galactinol synthase, a raffinosesynthetic enzyme; or a polynucleotide conferring resistance to tipburn,Fusarium oxysporum, Nasonovia ribisnigri, Sclerotinia sclerotiorum orother plant pathogens.

Foreign Protein Genes and Agronomic Genes

By means of the present invention, plants can be genetically engineeredto express various phenotypes of agronomic interest. Through thetransformation of celery, the expression of genes can be altered toenhance disease resistance, insect resistance, herbicide resistance,agronomic, nutritional quality, and other traits. Transformation canalso be used to insert DNA sequences which control or help controlmale-sterility. DNA sequences native to celery, as well as non-nativeDNA sequences, can be transformed into celery and used to alter levelsof native or non-native proteins. Various promoters, targetingsequences, enhancing sequences, and other DNA sequences can be insertedinto the genome for the purpose of altering the expression of proteins.Reduction of the activity of specific genes (also known as genesilencing or gene suppression) is desirable for several aspects ofgenetic engineering in plants.

Many techniques for gene silencing are well known to one of skill in theart, including, but not limited to, knock-outs (such as by insertion ofa transposable element such as mu (Vicki Chandler, The Maize Handbook,Ch. 118 (Springer-Verlag 1994)) or other genetic elements such as a FRTand Lox that are used for site specific integrations, antisensetechnology (see, e.g., Sheehy, et al., PNAS USA, 85:8805-8809 (1988);and U.S. Pat. Nos. 5,107,065, 5,453,566, and 5,759,829); co-suppression(e.g., Taylor, Plant Cell, 9:1245 (1997); Jorgensen, Trends Biotech.,8(12):340-344 (1990); Flavell, PNAS USA, 91:3490-3496 (1994); Finnegan,et al., Bio/Technology, 12:883-888 (1994); Neuhuber, et al., Mol. Gen.Genet., 244:230-241 (1994)); RNA interference (Napoli, et al., PlantCell, 2:279-289 (1990); U.S. Pat. No. 5,034,323; Sharp, Genes Dev.,13:139-141 (1999); Zamore, et al., Cell, 101:25-33 (2000); Montgomery,et al., PNAS USA, 95:15502-15507 (1998)), virus-induced gene silencing(Burton, et al., Plant Cell, 12:691-705 (2000); Baulcombe, Curr. Op.Plant Bio., 2:109-113 (1999)); target-RNA-specific ribozymes (Haseloff,et al., Nature, 334: 585-591 (1988)); hairpin structures (Smith, et al.,Nature, 407:319-320 (2000); WO 99/53050; WO 98/53083); MicroRNA(Aukerman & Sakai, Plant Cell, 15:2730-2741 (2003)); ribozymes(Steinecke, et al., EMBO J., 11:1525 (1992); Perriman, et al., AntisenseRes. Dev., 3:253 (1993)); oligonucleotide mediated targeted modification(e.g., WO 03/076574 and WO 99/25853); Zn-finger targeted molecules(e.g., WO 01/52620, WO 03/048345, and WO 00/42219); and other methods orcombinations of the above methods known to those of skill in the art.

Likewise, by means of the present invention, agronomic genes can beexpressed in transformed plants. More particularly, plants can begenetically engineered to express various phenotypes of agronomicinterest. Exemplary nucleotide sequences and/or native loci that conferat least one trait of interest, which optionally may be conferred oraltered by genetic engineering, transformation or introgression of atransformed event include, but are not limited to, those categorizedbelow:

1. Genes that Confer Resistance to Pests or Disease and that Encode:

A. Plant disease resistance genes. Plant defenses are often activated byspecific interaction between the product of a disease resistance gene(R) in the plant and the product of a corresponding avirulence (Avr)gene in the pathogen. A plant line can be transformed with a clonedresistance gene to engineer plants that are resistant to specificpathogen strains. See, for example Jones et al., Science 266:789 (1994)(cloning of the tomato Cf-9 gene for resistance to Cladosporium fulvum);Martin et al., Science 262:1432 (1993) (tomato Pto gene for resistanceto Pseudomonas syringae pv. Tomato encodes a protein kinase); Mindrinoset al., Cell 78:1089 (1994) (Arabidopsis RSP2 gene for resistance toPseudomonas syringae).

B. A Bacillus thuringiensis protein, a derivative thereof or a syntheticpolypeptide modeled thereon. See, for example, Geiser et al., Gene48:109 (1986), who disclose the cloning and nucleotide sequence of a Btδ-endotoxin gene. Moreover, DNA molecules encoding 6-endotoxin genes canbe purchased from American Type Culture Collection, Manassas, Virginia,for example, under ATCC Accession Nos. 40098, 67136, 31995 and 31998.

C. A lectin. See, for example, the disclosure by Van Damme et al., PlantMolec. Biol. 24:25 (1994), who disclose the nucleotide sequences ofseveral Clivia miniata mannose-binding lectin genes.

D. A vitamin-binding protein such as avidin. See PCT applicationUS93/06487, the contents of which are hereby incorporated by reference.The application teaches the use of avidin and avidin homologues aslarvicides against insect pests.

E. An enzyme inhibitor, for example, a protease or proteinase inhibitoror an amylase inhibitor. See, for example, Abe et al., J. Biol. Chem.262:16793 (1987) (nucleotide sequence of rice cysteine proteinaseinhibitor), Huub et al., Plant Molec. Biol. 21:985 (1993) (nucleotidesequence of cDNA encoding tobacco proteinase inhibitor I), Sumitani etal., Biosci. Biotech. Biochem. 57:1243 (1993) (nucleotide sequence ofStreptomyces nitrosporeus α-amylase inhibitor).

F. An insect-specific hormone or pheromone such as an ecdysteroid andjuvenile hormone, a variant thereof, a mimetic based thereon, or anantagonist or agonist thereof. See, for example, the disclosure byHammock et al., Nature 344:458 (1990), of baculovirus expression ofcloned juvenile hormone esterase, an inactivator of juvenile hormone.

G. An insect-specific peptide or neuropeptide which, upon expression,disrupts the physiology of the affected pest. For example, see thedisclosures of Regan, J. Biol. Chem. 269:9 (1994) (expression cloningyields DNA coding for insect diuretic hormone receptor), and Pratt etal., Biochem. Biophys. Res. Comm. 163:1243 (1989) (an allostatin isidentified in Diploptera puntata). See also U.S. Pat. No. 5,266,317 toTomalski et al., who disclose genes encoding insect-specific, paralyticneurotoxins.

H. An insect-specific venom produced in nature by a snake, a wasp, etc.For example, see Pang et al., Gene 116:165 (1992), for disclosure ofheterologous expression in plants of a gene coding for a scorpioninsectotoxic peptide.

I. An enzyme responsible for a hyper accumulation of a monoterpene, asesquiterpene, a steroid, hydroxamic acid, a phenylpropanoid derivativeor another non-protein molecule with insecticidal activity.

J. An enzyme involved in the modification, including thepost-translational modification, of a biologically active molecule; forexample, a glycolytic enzyme, a proteolytic enzyme, a lipolytic enzyme,a nuclease, a cyclase, a transaminase, an esterase, a hydrolase, aphosphatase, a kinase, a phosphorylase, a polymerase, an elastase, achitinase and a glucanase, whether natural or synthetic. See PCTapplication WO 93/02197 in the name of Scott et al., which discloses thenucleotide sequence of a callase gene. DNA molecules which containchitinase-encoding sequences can be obtained, for example, from ATCCunder Accession Nos. 39637 and 67152. See also Kramer et al., InsectBiochem. Molec. Biol. 23:691 (1993), who teach the nucleotide sequenceof a cDNA encoding tobacco hookworm chitinase, and Kawalleck et al.,Plant Molec. Biol. 21:673 (1993), who provide the nucleotide sequence ofthe parsley ubi4-2 polyubiquitin gene.

K. A molecule that stimulates signal transduction. For example, see thedisclosure by Botella et al., Plant Molec. Biol. 24:757 (1994), ofnucleotide sequences for mung bean calmodulin cDNA clones, and Griess etal., Plant Physiol. 104:1467 (1994), who provide the nucleotide sequenceof a maize calmodulin cDNA clone.

L. A hydrophobic moment peptide. See PCT application WO 95/16776(disclosure of peptide derivatives of Tachyplesin which inhibit fungalplant pathogens) and PCT application WO 95/18855 (teaches syntheticantimicrobial peptides that confer disease resistance), the respectivecontents of which are hereby incorporated by reference.

M. A membrane permease, a channel former or a channel blocker. Forexample, see the disclosure of Jaynes et al., Plant Sci 89:43 (1993), ofheterologous expression of a cecropin-β, lytic peptide analog to rendertransgenic tobacco plants resistant to Pseudomonas solanacearum.

N. A viral-invasive protein or a complex toxin derived therefrom. Forexample, the accumulation of viral coat proteins in transformed plantcells imparts resistance to viral infection and/or disease developmenteffected by the virus from which the coat protein gene is derived, aswell as by related viruses. See Beachy et al., Ann. rev. Phytopathol.28:451 (1990). Coat protein-mediated resistance has been conferred upontransformed plants against alfalfa mosaic virus, cucumber mosaic virus,tobacco streak virus, potato virus X, potato virus Y, tobacco etchvirus, tobacco rattle virus and tobacco mosaic virus. Id.

O. An insect-specific antibody or an immunotoxin derived therefrom.Thus, an antibody targeted to a critical metabolic function in theinsect gut would inactivate an affected enzyme, killing the insect. Cf.Taylor et al., Abstract #497, Seventh Int'l Symposium on MolecularPlant-Microbe Interactions (Edinburgh, Scotland) (1994) (enzymaticinactivation in transgenic tobacco via production of single-chainantibody fragments).

P. A virus-specific antibody. See, for example, Tavladoraki et al.,Nature 366:469 (1993), who show that transgenic plants expressingrecombinant antibody genes are protected from virus attack.

Q. A developmental-arrestive protein produced in nature by a pathogen ora parasite. Thus, fungal endo α-1, 4-D-polygalacturonases facilitatefungal colonization and plant nutrient release by solubilizing plantcell wall homo-α-1,4-D-galacturonase. See Lamb et al., Bio/Technology10:1436 (1992). The cloning and characterization of a gene which encodesa celery endopolygalacturonase-inhibiting protein is described byToubart et al., Plant 2:367 (1992).

R. A developmental-arrestive protein produced in nature by a plant. Forexample, Logemann et al., Bio/Technology 10:305 (1992), have shown thattransgenic plants expressing the barley ribosome-inactivating gene havean increased resistance to fungal disease.

S. A lettuce mosaic potyvirus (LMV) coat protein gene introduced intocelery in order to increase its resistance to LMV infection. See Dinantet al., Molecular Breeding. 1997, 3: 1, 75-86.

T. Genes involved in the Systemic Acquired Resistance (SAR) Responseand/or the pathogenesis-related genes. Briggs, S., Current Biology, 5(2)(1995).

U. Antifungal genes. See Cornelissen and Melchers, Plant Physiol.,101:709-712 (1993); Parijs et al., Planta 183:258-264 (1991) andBushnell et al., Can. J. of Plant Path. 20(2):137-149 (1998).

V. Genes that confer resistance to Phytophthora root rot, such as theRps 1, Rps 1-a, Rps 1-b, Rps 1-c, Rps 1-d, Rps 1-e, Rps 1-k, Rps 2, Rps3-a, Rps 3-b, Rps 3-c, Rps 4, Rps 5, Rps 6, Rps 7 and other Rps genes.See, for example, Shoemaker et al., Phytophthora Root Rot ResistanceGene Mapping in Soybean, Plant Genome IV Conference, San Diego, Calif.(1995).

2. Genes that Confer Resistance to an Herbicide:

A. An herbicide that inhibits the growing point or meristem, such as animidazolinone or a sulfonylurea. Exemplary genes in this category codefor mutant ALS and AHAS enzyme as described, for example, by Lee et al.,EMBO J. 7:1241 (1988), and Miki et al., Theor. Appl. Genet. 80:449(1990), respectively.

B. Glyphosate (resistance impaired by mutant5-enolpyruvlshikimate-3-phosphate synthase (EPSPS) and aroA genes,respectively) and other phosphono compounds such as glufosinate(phosphinothricin acetyl transferase, PAT and Streptomyces hygroscopicusphosphinothricin-acetyl transferase, bar, genes), and pyridinoxy orphenoxy proprionic acids and cyclohexones (ACCase inhibitor-encodinggenes). See, for example, U.S. Pat. No. 4,940,835 to Shah, et al., whichdiscloses the nucleotide sequence of a form of EPSPs which can conferglyphosate resistance. A DNA molecule encoding a mutant aroA gene can beobtained under ATCC accession number 39256, and the nucleotide sequenceof the mutant gene is disclosed in U.S. Pat. No. 4,769,061 to Comai. Seealso Umaballava-Mobapathie in Transgenic Research. 1999, 8: 1, 33-44that discloses Lactuca sativa resistant to glufosinate. European patentapplication No. 0 333 033 to Kumada et al., and U.S. Pat. No. 4,975,374to Goodman et al., disclose nucleotide sequences of glutamine synthetasegenes which confer resistance to herbicides such as L-phosphinothricin.The nucleotide sequence of a phosphinothricin-acetyl-transferase gene isprovided in European application No. 0 242 246 to Leemans et al.,DeGreef et al., Bio/Technology 7:61 (1989), describe the production oftransgenic plants that express chimeric bar genes coding forphosphinothricin acetyl transferase activity. Exemplary of genesconferring resistance to phenoxy proprionic acids and cyclohexones, suchas sethoxydim and haloxyfop are the Acc1-S1, Acc1-S2 and Acc1-S3 genesdescribed by Marshall et al., Theor. Appl. Genet. 83:435 (1992).

C. An herbicide that inhibits photosynthesis, such as a triazine (psbAand gs+ genes) and a benzonitrile (nitrilase gene). Przibilla et al.,Plant Cell 3:169 (1991), describe the transformation of Chlamydomonaswith plasmids encoding mutant psbA genes. Nucleotide sequences fornitrilase genes are disclosed in U.S. Pat. No. 4,810,648 to Stalker, andDNA molecules containing these genes are available under ATCC AccessionNos. 53435, 67441, and 67442. Cloning and expression of DNA coding for aglutathione S-transferase is described by Hayes et al., Biochem. J.285:173 (1992).

D. Acetohydroxy acid synthase, which has been found to make plants thatexpress this enzyme resistant to multiple types of herbicides, has beenintroduced into a variety of plants. See Hattori et al., Mol. Gen.Genet. 246:419, 1995. Other genes that confer tolerance to herbicidesinclude a gene encoding a chimeric protein of rat cytochrome P4507A1 andyeast NADPH-cytochrome P450 oxidoreductase (Shiota et al., PlantPhysiol., 106:17, 1994), genes for glutathione reductase and superoxidedismutase (Aono et al., Plant Cell Physiol. 36:1687, 1995), and genesfor various phosphotransferases (Datta et al., Plant Mol. Biol. 20:619,1992).

E. Protoporphyrinogen oxidase (PPO; protox) is the target of thePPO-inhibitor class of herbicides; a PPO-inhibitor resistant PPO genewas recently identified in Amaranthus tuberculatus (Patzoldt et al.,PNAS, 103(33):12329-2334, 2006). PPO is necessary for the production ofchlorophyll, which is necessary for all plant survival. The protoxenzyme serves as the target for a variety of herbicidal compounds. Theseherbicides also inhibit growth of all the different species of plantspresent, causing their total destruction. The development of plantscontaining altered protox activity which are resistant to theseherbicides are described in U.S. Pat. Nos. 6,288,306, 6,282,837,5,767,373, and International Publication WO 01/12825.

F. Genes that confer resistance to auxin or synthetic auxin herbicides.For example an aryloxyalkanoate dioxygenase (AAD) gene may conferresistance to arlyoxyalkanoate herbicides, such as 2,4-D, as well aspyridyloxyacetate herbicides, such as described in U.S. Pat. No.8,283,522, and US2013/0035233. In other examples, a dicambamonooxygenase (DMO) is used to confer resistance to dicamba. Otherpolynucleotides of interest related to auxin herbicides and/or usesthereof include, for example, the descriptions found in U.S. Pat. Nos.8,119,380; 7,812,224; 7,884,262; 7,855,326; 7,939,721; 7,105,724;7,022,896; 8,207,092; US2011/067134; and US2010/0279866. Any of theabove listed herbicide genes (1-6) can be introduced into the claimedcelery cultivar through a variety of means including, but not limitedto, transformation and crossing.

3. Genes that Confer or Contribute to a Value-Added Trait, Such as:

A. Increased iron content of the celery, for example by transforming aplant with a soybean ferritin gene as described in Goto et al., ActaHorticulturae. 2000, 521, 101-109.

B. Decreased nitrate content of leaves, for example by transforming acelery with a gene coding for a nitrate reductase. See for exampleCurtis et al., Plant Cell Report. 1999, 18:11, 889-896.

C. Increased sweetness of the celery by transferring a gene coding formonellin that elicits a flavor 100,000 times sweeter than sugar on amolar basis. See Penarrubia et al., Biotechnology. 1992, 10:5, 561-564.

D. Modified fatty acid metabolism, for example, by transforming a plantwith an antisense gene of stearyl-ACP desaturase to increase stearicacid content of the plant. See Knultzon et al., Proc. Natl. Acad. Sci.USA 89:2625 (1992).

E. Modified carbohydrate composition effected, for example, bytransforming plants with a gene coding for an enzyme that alters thebranching pattern of starch. See Shiroza et al., J. Bacteriol. 170:810(1988) (nucleotide sequence of Streptococcus mutantsfructosyltransferase gene), Steinmetz et al., Mol. Gen. Genet. 20:220(1985) (nucleotide sequence of Bacillus subtilis levansucrase gene), Penet al., Bio/Technology 10:292 (1992) (production of transgenic plantsthat express Bacillus licheniformis α-amylase), Elliot et al., PlantMolec. Biol. 21:515 (1993) (nucleotide sequences of tomato invertasegenes), Søgaard et al., J. Biol. Chem. 268:22480 (1993) (site-directedmutagenesis of barley α-amylase gene), and Fisher et al., Plant Physiol.102:1045 (1993) (maize endosperm starch branching enzyme II).

F. Altered antioxidant content or composition, such as alteration oftocopherol or tocotrienols. See, for example, U.S. Pat. Nos. 6,787,683,7,154,029, WO 00/68393 (involving the manipulation of antioxidant levelsthrough alteration of a phytl prenyl transferase (ppt)); WO 03/082899(through alteration of a homogentisate geranyl geranyl transferase(hggt)).

G. Modified bolting tolerance in plants for example, by transferring agene encoding for gibberellin 2-oxidase (U.S. Pat. No. 7,262,340).Bolting has also been modified using non-transformation methods; seeWittwer, S. H., et al. (1957) Science. 126(3262): 30-31; Booij, R. etal., (1995) Scientia Horticulturae. 63:143-154; and Booij, R. et al.,(1994) Scientia Horticulturae. 58:271-282.

H. Decreased browning of the celery, for example by transforming a plantwith an siRNA, RNAi or microRNA vector, or other suppression sequencecoding for polyphenol oxidase (PPO) to silence the expression of PPOgenes. See Araji et al. (2014) Plant Physiology 164:1191-1203, Chi etal. (2014) BMC Plant Biology 14:62, and Carter, N., (2012) Petition forDetermination of Nonregulated Status: Arctic™ Apple (Malta×domestica)Events GD743 and GS784, received by APHIS.

4. Genes that Control Male-Sterility

A. Introduction of a deacetylase gene under the control of atapetum-specific promoter and with the application of the chemicalN—Ac-PPT. See international publication WO 01/29237.

B. Introduction of various stamen-specific promoters. See internationalpublications WO 92/13956 and WO 92/13957.

C. Introduction of the barnase and the barstar genes. See Paul et al.,Plant Mol. Biol. 19:611-622, 1992).

For additional examples of nuclear male and female sterility systems andgenes, see also, U.S. Pat. Nos. 5,859,341, 6,297,426, 5,478,369,5,824,524, 5,850,014, and 6,265,640, all of which are herebyincorporated by reference.

5. Genes that Affect Abiotic Stress Resistance

Genes that affect abiotic stress resistance (including but not limitedto flowering, seed development, enhancement of nitrogen utilizationefficiency, altered nitrogen responsiveness, drought resistance ortolerance, cold resistance or tolerance, high or low light intensity,and salt resistance or tolerance) and increased yield under stress. Forexample, see: WO 00/73475 where water use efficiency is altered throughalteration of malate; U.S. Pat. Nos. 5,892,009, 5,965,705, 5,929,305,5,891,859, 6,417,428, 6,664,446, 6,706,866, 6,717,034, 6,801,104, WO2000/060089, WO 2001/026459, WO 2001/035725, WO 2001/034726, WO2001/035727, WO 2001/036444, WO 2001/036597, WO 2001/036598, WO2002/015675, WO 2002/017430, WO 2002/077185, WO 2002/079403, WO2003/013227, WO 2003/013228, WO 2003/014327, WO 2004/031349, WO2004/076638, WO 98/09521, and WO 99/38977 describing genes, includingCBF genes and transcription factors effective in mitigating the negativeeffects of freezing, high salinity, and drought on plants, as well asconferring other positive effects on plant phenotype; U.S. Publ. No.2004/0148654 and WO 01/36596, where abscisic acid is altered in plantsresulting in improved plant phenotype, such as increased yield and/orincreased tolerance to abiotic stress; WO 2000/006341, WO 04/090143,U.S. Pat. Nos. 7,531,723 and 6,992,237, where cytokinin expression ismodified resulting in plants with increased stress tolerance, such asdrought tolerance, and/or increased yield. See also, WO 02/02776, WO2003/052063, JP 2002281975, U.S. Pat. No. 6,084,153, WO 01/64898, andU.S. Pat. Nos. 6,177,275 and 6,107,547 (enhancement of nitrogenutilization and altered nitrogen responsiveness). For ethylenealteration, see, U.S. Publ. Nos. 2004/0128719, 2003/0166197, and WO2000/32761. For plant transcription factors or transcriptionalregulators of abiotic stress, see, e.g., U.S. Publ. Nos. 2004/0098764 or2004/0078852.

Other genes and transcription factors that affect plant growth andagronomic traits, such as yield, flowering, plant growth, and/or plantstructure, can be introduced or introgressed into plants. See, e.g., WO97/49811 (LHY), WO 98/56918 (ESD4), WO 97/10339, U.S. Pat. No. 6,573,430(TFL), 6,713,663 (FT), 6,794,560, 6,307,126 (GAI), WO 96/14414 (CON), WO96/38560, WO 01/21822 (VRN1), WO 00/44918 (VRN2), WO 99/49064 (GI), WO00/46358 (FRI), WO 97/29123, WO 99/09174 (D8 and Rht), WO 2004/076638,and WO 004/031349 (transcription factors).

Tissue Culture

Further reproduction of the variety can occur by tissue culture andregeneration. Tissue culture of various tissues of celery andregeneration of plants there from is well known and widely published.For example, reference may be had to Teng et al., Hort Science. 1992,27: 9, 1030-1032 Teng et al., Hort Science. 1993, 28: 6, 669-1671, Zhanget al., Journal of Genetics and Breeding. 1992, 46: 3, 287-290, Webb etal., Plant Cell Tissue and Organ Culture. 1994, 38: 1, 77-79, Curtis etal., Journal of Experimental Botany. 1994, 45: 279, 1441-1449, Nagata etal., Journal for the American Society for Horticultural Science. 2000,125: 6, 669-672, and Ibrahim et al., Plant Cell, Tissue and OrganCulture. (1992), 28(2): 139-145. It is clear from the literature thatthe state of the art is such that these methods of obtaining plants areroutinely used and have a very high rate of success. Thus, anotheraspect of this invention is to provide cells which upon growth anddifferentiation produce celery plants having the physiological andmorphological characteristics of variety TBG 45.

As used herein, the term “tissue culture” indicates a compositioncomprising isolated cells of the same or a different type or acollection of such cells organized into parts of a plant. Exemplarytypes of tissue cultures are protoplasts, calli, plant clumps, and plantcells that can generate tissue culture that are intact in plants orparts of plants, such as embryos, hypocotyls, pollen, flowers, seeds,leaves, stems, roots, root tips, pistils, anthers, meristematic cellsand the like. Means for preparing and maintaining plant tissue cultureare well known in the art. By way of example, a tissue culturecomprising organs has been used to produce regenerated plants. U.S. Pat.Nos. 5,959,185; 5,973,234 and 5,977,445 describe certain techniques, thedisclosures of which are incorporated herein by reference.

Industrial Uses of Celery Cultivar TBG 45

Celery may be used in a variety of manners including but not limited to,use in salads, soups, being filled with cheese, soybean, vegetable,peanut butter, or dairy type products, served raw, cooked, baked orfrozen, served as sticks, pieces, diced, or dipped like potato chips, orused as straws.

Tables

In the tables that follow, the traits and characteristics of celerycultivar TBG 45 are given compared to other celery cultivars. Colorreferences made in the Tables refer to the Munsell Color Chart.

Tables 2A and 2B show the results of a trial comparing characteristicsof celery cultivar TBG 45 to celery varieties TBG 43, TBG 29 (U.S. Pat.No. 10,440,913), ADS-1 (U.S. Pat. No. 6,822,143), TBG 34 (U.S. Pat. No.10,306,857), TBG 37, TBG 28, TBG 33 (U.S. Pat. No. 9,713,308), ADS-22(U.S. Pat. No. 8,598,424), Hill's Special (U.S. PVP Certificate No.009500019), ADS-20 (U.S. Pat. No. 8,524,993), Challenger, Sonora (U.S.PVP Certificate No. 009900063), Conquistador, Command (U.S. Pat. No.7,193,142; U.S. PVP Certificate No. 200400326), Mission, Floribelle,ADS-23 (U.S. Pat. No. 8,598,425), TBG 31 (U.S. Pat. No. 9,439,374), TBG54, TBG 35, TBG 32 (U.S. Pat. No. 9,433,162), Tall Utah 52-70 ‘R’Strain, and Tall Utah 52-75. The trial was transplanted in Oxnard,California on Aug. 6, 2020, and evaluated on Nov. 5, 2020 (91 days).This trial was grown in a production block that has fairly high levelsof Fusarium oxysporum race 2. Under these conditions varieties wereevaluated for relative tolerance to the disease. The plant population(58,080 plants to the acre) was higher than the commercial norm ofapproximately 45,000 to 47,000 plants to the acre. Table 2A, column 1shows the characteristic and columns 2-13 show the results for TBG 45,TBG 43, TBG 29, ADS-1, TBG 34, TBG 37, TBG 28, TBG 33, ADS-22, Hill'sSpecial, ADS-20, and Challenger, respectively. Table 2B, column 1 showsthe characteristic and columns 2-13 show the results for Sonora,Conquistador, Command, Mission, Floribelle, ADS-23, TBG 31, TBG 54, TBG35, TBG 32, Tall Utah 52-70 ‘R’ Strain, and Tall Utah 52-75,respectively. NA indicates data not available.

TABLE 2A TBG ADS- Hill's ADS- Chal- TBG 45 TBG 43 TBG 29 ADS-1 TBG 34TBG 37 TBG 28 33 22 Special 20 lenger Plant height Average 78.7 77.771.8 50.0 50.4 78.1 73.3 68.1 72.3 47.6 36.7 77.5 (cm) Range (72-84)(71-84) (67-74) (42-59) (44-54) (75-82) (57-87) (64-72) (62-82) (43-54)(28-46) (64-87) Whole plant Average  1.22  1.08  1.07 NA NA  1.06  0.38 0.76  0.81 NA NA  1.08 weight (kg) Range (0.79- (0.77- (0.77- (0.92-(0-1.4) (0.19- (0.67- (0.41- 1.56) 1.37) 1.33) 1.49) 1.2) 1.07) 1.61)Trimmed plant Average  0.93  0.85  0.92 NA NA  0.84  0.29  0.71  0.69 NANA  0.89 weight Range (0.64- (0.59- (0.63- (0.71- (0-1.03) (0.38- (0.57-(0.63- (kg @ 35.6 cm) 1.14) 1.04) 1.1) 1.11) 0.99) 0.9) 1.23) Number ofAverage 11.7 11.1 12.3 NA NA 11.1  3.3 11.6 10.2 NA NA 10.5 outerpetioles Range (8-14) (8-13) (10-14) (9-12) (0-13) (9-14) (9-12) (8-13)(>35.6 cm) Number of Average  4.6  5.0  5.5 NA NA  5.2  1.8  4.4  6.3 NANA  7.4 inner petioles Range (4-6) (4-6) (5-6) (4-6) (0-7) (3-6) (4-10)(5-9) (<35.6 cm) Length of outer Average 30.1 27.3 25.1 17.6 23.0 29.727.5 23.3 21.6 21.3 16.0 29.2 petioles to the Range (28-32) (24-29.3)(23.3-28) (14.3- (18.3- (26.7- (19-33) (21-25) (21- (17.3- (13-20)(23.7- joint (cm) 21) 27.7) 33) 22.3) 25.3) 34.7) Width of outer Average22.8 22.5 23.3 NA NA 21.7  7.1 19.7 23.1 NA NA 20.9 petioles at theRange (20-24.7) (19.7- (20.7-25) (19-25) (0-25) (16.7- (21- (14.7-midrib (mm) 25.3) 21.3) 24.7) 24.3) Thickness of Average 10.7 11.5 11.1NA NA 10.9  3.8  9.9  9.4 NA NA 10.4 outer petioles Range (9.3-(9.3-13.3) (9.7-13) (10.3- (0-13.3) (8.7- (7.7- (5-12.7) at the midrib11.3) 12) 10.7) 10.3) (mm) Petiole color 5gy 7/8 5gy 7/6 5gy 7/4 NA NA5gy 7/6 5gy 7/6 5gy 5gy NA NA 5gy (Munsell color) 7/6 7/6 7/6 Leaf color5gy 3/4 5gy 3/4 5gy 3/4 NA NA 5gy 3/4 5gy 3/4 5gy 5gy NA NA 5gy (Munsellcolor) 3/4 3/4 3/4 Petiole Slight Smooth Slight NA NA Smooth SmoothSlight Smooth NA NA Smooth smoothness rib/Rib rib/Rib rib/Rib Petiolecup Cup Cup Cup NA NA Cup Cup Cup Cup NA NA Cup/ Deep cup % Marketable100% 100% 100% 0% 0% 100% 30% 70% 100% 0% 0%  70% Disease: OverallAverage  5  5  5  2.5  2.5  5  4.5  4  5  3  2  4 fusarium Range  5  5 5 2-3 2-3  5 3-5 3-5  5 2-3 1-2 3-5 ratings (0 = dead, 5 = tolerant)Defects: % Top pith  0%  0%  0% NA NA  0%  0%  0%  0% NA NA  10% % Buttpith  0%  0%  0% NA NA  0%  0%  0%  0% NA NA  10% % Butt crack  0%  0% 0% NA NA  0% 50%  0%  0% NA NA  30% % Twist  0%  0%  0% NA NA  0% 70% 0%  0% NA NA 100% % Black heart  0%  0%  0% NA NA  0%  0%  0%  0% NA NA 10%

TABLE 2B Tall Utah 52-70 Tall Conquis- ‘R’ Utah Sonora tador CommandMission Floribelle ADS-23 TBG 31 TBG 54 TBG 35 TBG 32 Strain 52-75 Plantheight Average 21.0 32.1 51.3 32.9 12.3 41.6 37.6 26.1 23.8 29.9 17.024.5 (cm) Range (0-34) (0-49) (38-60) (0-45) (0-37) (0-60) (0-56) (0-45)(0-34) (0-49) (0-32) (0-37) Whole plant Average NA NA NA NA NA NA NA NANA NA NA NA weight (kg) Range Trimmed plant Average NA NA NA NA NA NA NANA NA NA NA NA weight Range (kg @ 35.6 cm) Number of Average NA NA NA NANA NA NA NA NA NA NA NA outer petioles Range (>35.6 cm) Number ofAverage NA NA NA NA NA NA NA NA NA NA NA NA inner petioles Range (<35.6cm) Length of outer Average 7.8 13.1 19.5 14.3 6.1 18.4 17.0 15.0 9.713.5 6.2 8.6 petioles to the Range (0-13) (0-21) (11-24.3) (0-21.7)(0-18.7) (0-26.7) (0-26) (0-27.7) (0-14.3) (0-23) (0-12.7) (0-14) joint(cm) Width of outer Average NA NA NA NA NA NA NA NA NA NA NA NA petiolesat the Range midrib (mm) Thickness of Average NA NA NA NA NA NA NA NA NANA NA NA outer petioles Range at the midrib (mm) Petiole color NA NA NANA NA NA NA NA NA NA NA NA (Munsell color) Leaf color NA NA NA NA NA NANA NA NA NA NA NA (Munsell color) Petiole smoothness NA NA NA NA NA NANA NA NA NA NA NA Petiole cup NA NA NA NA NA NA NA NA NA NA NA NA %Marketable 0% 0% 0% 0% 0% 0% 0% 0% 0% 0% 0% 0% Disease: Overall Average1 1 3 1.5 1 1 2.5 1 1 2 1 1 fusarium Range 0-2 0-2 2-3 0-2 0-2 0-2 0-30-2 0-1 0-3 0-1 0-2 ratings (0 = dead, 5 = tolerant) Defects: % Top pithNA NA NA NA NA NA NA NA NA NA NA NA % Butt pith NA NA NA NA NA NA NA NANA NA NA NA % Butt crack NA NA NA NA NA NA NA NA NA NA NA NA % Twist NANA NA NA NA NA NA NA NA NA NA NA % Black heart NA NA NA NA NA NA NA NANA NA NA NA

As shown in Tables 2A and 2B, celery cultivars TBG 45, TBG 43, TBG 29,and TBG 37 appeared to be free from race 2 infection while all othervarieties expressed various levels of susceptibility (fusarium levels inthe root and overall performance ratings). TBG 45, TBG 43, TBG 29, andTBG 37 showed the best tolerance to Fusarium oxysporum race 2 whenoverall infection ratings and percent marketability were considered.These were followed by TBG 28 and Challenger which also had other majordefects that impacted marketability; some as a result of the fusariuminfection. TBG 45 and TBG 29 looked most similar when comparing trimplant, although TBG 45 had more initial untrimmed weight which may havebeen due to its greater plant height and length to first joint whichwere more similar to TBG 37. However, TBG 37, TBG 43, and TBG 29 werevery similar in slightly lower trimmed weight. TBG 45 and TBG 29 werealso more similar for ribbiness. TBG 45 produced longer petioles thanTBG 43 and TBG 29.

Tables 3A and 3B show the results of a trial comparing characteristicsof celery cultivar TBG 45 to celery varieties TBG 43, TBG 29, ADS-1, TBG34, TBG 37, TBG 28, TBG 33, ADS-22, Hill's Special, ADS-20, Challenger,Sonora, Conquistador, Command, Mission, ADS-23, TBG 31, TBG 54, TBG 35,TBG 32, TBG 53, Tall Utah 52-70 ‘R’ Strain, and Tall Utah 52-75. Thetrial was transplanted in Oxnard, California on Aug. 9, 2020, andevaluated on Dec. 12, 2020 (125 days) in a block that has been developedwith especially high levels of Fusarium oxysporum race 2 and slight tomoderate levels from Fusarium oxysporum race 4. The plant population(58,080 plants to the acre) was higher than the commercial norm ofapproximately 45,000 to 47,000 plants to the acre. Table 3A, column 1shows the characteristic and columns 2-13 show the results for TBG 45,TBG 43, TBG 29, ADS-1, TBG 34, TBG 37, TBG 28, TBG 33, ADS-22, Hill'sSpecial, ADS-20, and Challenger, respectively. Table 3B, column 1 showsthe characteristic and columns 2-13 show the results for Sonora,Conquistador, Command, Mission, ADS-23, TBG 31, TBG 54, TBG 35, TBG 32,TBG 53, Tall Utah 52-70 ‘R’ Strain, and Tall Utah 52-75, respectively.NA indicates data not available.

TABLE 3A Hill's TBG 45 TBG 43 TBG 29 ADS-1 TBG 34 TBG 37 TBG 28 TBG 33ADS-22 Special ADS-20 Challenger Plant height Average 83.1 84.6 60.4 9.921.2 37.3 6.4 4.1 16.5 7.7 10.5 24.4 (cm) Range (80-89) (78-93) (0-81)(0-27) (0-32) (0-58) (0-36) (0-27) (0-63) (0-35) (0-33) (0-56) Wholeplant Average 1.24 1.04 0.74 NA NA NA NA NA NA NA NA NA weight (kg)Range (0.94-1.53) (0.53-1.33) (0-1.41) Trimmed plant Average 0.91 0.750.58 NA NA NA NA NA NA NA NA NA weight Range (0.71-1.11) (0.37-1)(0-1.09) (kg @ 35.6 cm) Number of Average 12.6 11.1 7.2 NA NA NA NA NANA NA NA NA outer petioles Range (11-14) (9-14) (0-11) (>35.6 cm) Numberof Average 4.3 4.4 3.3 NA NA NA NA NA NA NA NA NA inner petioles Range(3-6) (3-5) (0-5) (<35.6 cm) Length of outer Average 32.1 26.6 20.0 3.88.1 15.9 2.2 1.3 5.3 2.6 4.1 8.4 petioles to the Range (26.7-38(21.3-29) (0-27) (0-12) (0-14) (0-25) (0-12) (0-8) (0-22) (0-12) (0-14)(0-21) joint (cm) Width of outer Average 23.0 21.7 16.3 NA NA NA NA NANA NA NA NA petioles at the Range (22-24.7) (17.3-24.7) (0-26) midrib(mm) Thickness of Average 11.0 11.9 9.1 NA NA NA NA NA NA NA NA NA outerpetioles Range (10-12.7) (9.7-13.3) (0-13.7) at the midrib (mm) Petiolesmoothness Smooth/ Smooth Smooth/ NA NA NA NA NA NA NA NA NA Slight ribSlight rib Petiole cup Slight Cup Cup NA NA NA NA NA NA NA NA NA cup/Cup% Marketable 100% 80% 30% NA NA NA NA NA NA NA NA NA Disease: Survivalrate Healthy 100% 80% 30%  0%  0%  0%  0%  0%  0%  0%  0% 10% (%) Dying 0% 20% 50% 47% 80% 80% 35% 27% 30% 32% 50% 50% Dead  0%  0% 20% 53% 20%20% 65% 73% 70% 68% 50% 40% Overall Average 5 4.8 3 0.9 1.9 3.2 0.6 0.51.1 0.7 1.2 1.8 fusarium Range (5-5) (3-5) (0-4) (0-3) (0-3) (0-5) (0-2)(0-3) (0-4) (0-2) (0-3) (0-3) ratings (0 = dead, 5 = tolerant) Rootfusarium Average 5 3.6 3 0.7 1.1 0.8 0.4 0.3 0.4 0.5 0.7 1.2 ratingsRange (4-5) (1-5) (0-4) (0-2) (0-2) (0-1) (0-1) (0-1) (0-2) (0-1) (0-2)(0-2) (0 = dead, 5 = tolerant)

TABLE 3B Tall Utah 52-70 Tall ′R′ Utah Sonora Conquistador CommandMission ADS-23 TBG 31 TBG 54 TBG 35 TBG 32 TBG 53 Strain 52-75 Plantheight Average 12.6 15.4 12.8 14.0 0.0 1.5 5.0 0.6 3.4 3.0 1.7 1.7 (cm)Range (0-33) (0-37) (0-38) (0-30) (0-0) (0-13) (0-22) (0-12) (0-23)(0-20) (0-19) (0-22) Whole plant Average NA NA NA NA NA NA NA NA NA NANA NA weight (kg) Range Trimmed plant Average NA NA NA NA NA NA NA NA NANA NA NA weight Range (kg @ 35.6 cm) Number of Average NA NA NA NA NA NANA NA NA NA NA NA outer petioles Range (>35.6 cm) Number of Average NANA NA NA NA NA NA NA NA NA NA NA inner petioles Range (<35.6 cm) Lengthof outer Average 4.8 4.1 3.7 4.6 0.0 0.6 2.8 0.3 1.2 1.3 0.8 0.7petioles to the Range (0-14) (0-14) (0-12) (0-10) (0-0) (0-8) (0-8)(0-6) (0-9) (0-8) (0-10) (0-8) joint (cm) Width of outer Average NA NANA NA NA NA NA NA NA NA NA NA petioles at the Range midrib (mm)Thickness of Average NA NA NA NA NA NA NA NA NA NA NA NA outer petiolesRange at the midrib (mm) Petiole smoothness NA NA NA NA NA NA NA NA NANA NA NA Petiole cup NA NA NA NA NA NA NA NA NA NA NA NA % Marketable NANA NA NA NA NA NA NA NA NA NA NA Disease: Survival rate Healthy  0%  0% 0%  0%   0%  0%  0%  0%  0%  0%  0%  0% (%) Dying 48% 58% 45% 60%   0%12% 37%  5% 22% 22% 12% 12% Dead 52% 42% 55% 40% 100% 88% 63% 95% 78%78% 88% 88% Overall Average 0.8 1 0.9 1.1 0.0 0.2 0.4 0.1 0.4 0.5 0.30.3 fusarium Range (0-2) (0-2) (0-2) (0-2) (0-0) (0-1) (0-1) (0-1) (0-2)(0-1) (0-1) (0-2) ratings (0 = dead, 5 = tolerant) Root fusarium Average0.6 0.7 0.5 0.7 0.0 0.2 0.4 0.1 0.3 0.5 0.3 0.3 ratings Range (0-1)(0-2) (0-1) (0-1) (0-0) (0-1) (0-1) (0-1) (0-1) (0-1) (0-1) (0-1) (0 =dead, 5 = tolerant)

As shown in Tables 3A and 3B, under these conditions celery cultivar TBG45 demonstrated excellent tolerance to both Fusarium oxysporum race 2and race 4 as can be noted by fusarium ratings, marketability, andsurvivable rates. TBG 43 and TBG 29 were the next best performingvarieties in this trial. All three varieties are noted for tolerance torace 2, so the primary marketable differences between them in this trialis their tolerance level to race 4. Under these conditions TBG 45 beginsto show its true potential and value. While several varieties havetolerance to fusarium race 2 (Table 2), when fusarium race 4 is presentmany of these varieties falter. In this trial, TBG 45 was 100%marketable while TBG 43 was only 80% and TBG 29 was 30% marketable.Marketability also correlated very closely with the survival rates.

Tables 4-7 show the results of trials grown in a field in Oxnard,California that was created with especially high inoculum levels ofFusarium oxysporum race 4 in order to develop and evaluate cultivars fortolerance. This field has had two celery crops produced on it in everyyear since the disease was first documented in 2013. In fact, this isthe actual field where the disease was first observed. With no rotationor attempt to diminish inoculum levels, this field has gained inoculumlevels and infectious potential over the years and is currently one ofthe most heavily infested fields in existence.

Excluding seasonal and environmental variation these trials provided intime sequence from Table 4 through Table 7 demonstrate increasing levelsof infectious capability and the impact on many varieties. While most ofthe varieties in the trials are truly susceptible and essentially dead(% survivors, % marketable, etc.), celery cultivar TBG 45 hassurprisingly shown good levels of tolerance with all race 4 infectiousrates (Tables 4-7) throughout the trials (80% to 100% marketability).The next best variety, TBG 43, has performed poorer as residual fusariumrace 4 infectious capabilities have increased over time. While TBG 43has some tolerance capabilities, no other variety has shown similartolerances and especially none comparable to TBG 45.

Tables 4A and 4B show the results of a trial comparing characteristicsof celery cultivar TBG 45 to celery varieties TBG 43, TBG 29, ADS-1, TBG34, TBG 37, TBG 28, TBG 33, ADS-20, Challenger, Sonora, Conquistador,Mission, ADS-23, TBG 31, TBG 54, TBG 35, TBG 32, Tall Utah 52-70 ‘R’Strain, and Tall Utah 52-75. The trial was transplanted in Oxnard,California on Aug. 28, 2018, and evaluated on Jan. 13, 2019 (138 days).This trial was grown in a production block that has been developed withespecially high levels of Fusarium oxysporum race 4 levels in order toevaluate and develop varieties for increased tolerance to the disease.Not only is this field especially high in the disease, but it is alsothe location where the disease was first identified. The plantpopulation (58,080 plants to the acre) was higher than the commercialnorm of approximately 45,000 to 47,000 plants to the acre. Table 4A,column 1 shows the characteristic and columns 2-11 show the results forTBG 45, TBG 43, TBG 29, ADS-1, TBG 34, TBG 37, TBG 28, TBG 33, ADS-20,and Challenger, respectively. Table 4B, column 1 shows thecharacteristic and columns 2-11 show the results for Sonora,Conquistador, Mission, ADS-23, TBG 31, TBG 54, TBG 35, TBG 32, Tall Utah52-70 ‘R’ Strain, and Tall Utah 52-75, respectively. NA indicates datanot available.

TABLE 4A TBG 45 TBG 43 TBG 29 ADS-1 TBG 34 TBG 37 TBG 28 TBG 33 ADS-20Challenger Plant height (cm) Average 68.0 61.0 44.7 1.7 18.3 43.0 1.11.5 14.2 3.6 Range (64-73) (0-74) (0-66) (0-43) (0-58) (24-69) (0-30)(0-36) (0-47) (0-66) Whole plant Average 1.0 0.82 0.63 0.01 0.14 0.280.00 0.01 0.08 0.05 weight (kg) Range (0.62-1.34) (0-1.52) (0-1.42)(0-0.37) (0-0.86) (0.03-0.92) (0-0.1) (0-0.65) (0-0.48) (0-1.39) Trimmedplant Average 0.77 0.69 0.54 NA NA NA NA NA NA NA weight (kg) Range(0.46-1.07) (0-1.26) (0-1.25) Number of outer Average 11.10 9.3 6.0 NANA NA NA NA NA NA petioles Range (9-13) (0-14) (0-15) (>35.6 cm) Numberof inner Average 4.2 5.5 0.5 NA NA NA NA NA NA NA petioles Range (3-5)(0-8) (0-12) (<35.6 cm) Length of outer Average 28.7 20.5 15.9 NA 6.818.5 0.4 0.6 5.2 1.3 petioles to the Range (26.3-34) (0-28) (0-24)(0-23) (7-32) (0-10) (0-12) (0-17) (0-30) joint (cm) Width of outerAverage 23.0 19.7 13.1 NA NA NA NA NA NA NA petioles at the Range(19-25.3) (0-25) (0-25.7) midrib (mm) Thickness of Average 9.7 9.2 5.5NA NA NA NA NA NA NA outer petioles at Range (5.7-12.7) (0-12.3) (0-11)the midrib (mm) % Marketable 100% 70% 30% 0% 10%  0% 0% 0%  0% 0% %Survivors 100% 80% 85% 6% 40% 88% 5% 5% 42% 8%

TABLE 4B Tall Utah 52-70 ′R′ Tall Utah Sonora Conquistador MissionADS-23 TBG 31 TBG 54 TBG 35 TBG 32 Strain 52-75 Plant height (cm)Average 5.3 3.8 10.0 0.0 10.3 0.2 0.0 0.5 0.0 0.0 Range (0-47) (13-50)(0-52) (0-0) (0-57) (0-19) (0-0) (0-20) (0-0) (0-0) Whole plant Average0.05 0.03 0.00 0.0 0.07 0.00 0.0 0.00 0.00 0.00 weight (kg) Range(0-0.54) (0.02-0.6) (0-0) (0-0) (0-0.63) (0-0) (0-0) (0-0.08) (0-0)(0-0) Trimmed plant Average NA NA NA 0.0 NA NA 0.0 NA 0.0 0.0 weight(kg) Range (0-0) (0-0) (0-0) (0-0) Number of outer Average NA NA NA 0.0NA NA 0.0 NA 0.0 0.0 petioles Range (0-0) (0-0) (0-0) (0-0) (>35.6 cm)Number of inner Average NA NA NA 0.0 NA NA 0.0 NA 0.0 0.0 petioles Range(0-0) (0-0) (0-0) (0-0) (<35.6 cm) Length of outer Average 1.8 1.4 3.50.0 NA NA 0.0 0.2 0.0 0.0 petioles to the Range (0-16) (5-20) (0-17)(0-0) (0-0) (0-7) (0-0) (0-0) joint (cm) Width of outer Average NA NA NA0.0 NA NA 0.0 NA 0.0 0.0 petioles at the Range (0-0) (0-0) (0-0) (0-0)midrib (mm) Thickness of Average NA NA NA 0.0 NA NA 0.0 NA 0.0 0.0 outerpetioles at Range (0-0) (0-0) (0-0) (0-0) the midrib (mm) % Marketable 0%  0%  0% 0% 10% 0% 0% 0% 0% 0% % Survivors 15% 14% 25% 0% 27% 1% 0%3% 0% 0%

As shown in Tables 4A and 4B, under the fusarium conditions of thistrial, most varieties had a few to several plants that were still aliveat evaluation but were not able to sustain development of marketablecelery stalks. Most were barely alive and most of the data were notcollected for the non-marketable stalks (marked as NA=not available).Under these conditions celery cultivar TBG 45 was very tolerant (100%marketable), TBG 43 was moderately tolerant (70% marketable), and TBG 29produced 30% marketable stalks. No other varieties produced marketablestalks.

Tables 5A and 5B show the results of a trial comparing characteristicsof celery cultivar TBG 45 to celery varieties TBG 43, TBG 29, ADS-1, TBG34, TBG 37, TBG 28, TBG 33, ADS-22, ADS-20, Floribelle, ADS-23, TBG 31,TBG 54, ADS-2 (U.S. Pat. No. 7,365,248), ADS-8 (U.S. Pat. No. 6,812,385;U.S. PVP Certificate No. 200200021), TBG 35, TBG 32, and TBG 53. Thetrial was transplanted in Oxnard, California on Aug. 14, 2019, andevaluated on Dec. 23, 2019 (131 days). This trial was grown in aproduction block that has been developed with especially high levels ofFusarium oxysporum race 4 levels in order to evaluate and developvarieties for increased tolerance to the disease. Not only is this fieldespecially high in the disease, but it is also the location where thedisease was first identified. The plant population (58,080 plants to theacre) was higher than the commercial norm of approximately 45,000 to47,000 plants to the acre. Table 5A, column 1 shows the characteristicand columns 2-11 show the results for TBG 45, TBG 43, TBG 29, ADS-1, TBG34, TBG 37, TBG 28, TBG 33, ADS-22, and ADS-20, respectively. Table 5B,column 1 shows the characteristic and columns 2-10 show the results forFloribelle, ADS-23, TBG 31, TBG 54, ADS-2, ADS-8, TBG 35, TBG 32, andTBG 53, respectively. NA indicates data not available.

TABLE 5A TBG 45 TBG 43 TBG 29 ADS-1 TBG 34 TBG 37 TBG 28 TBG 33 ADS-22ADS-20 Plant height (cm) Average 80.10 32.70 NA NA NA NA NA NA NA NARange (0-85) (0-74) Whole plant Average 1.35 0.65 NA NA NA NA NA NA NANA weight (kg) Range (0-1.5) (0-1.42) Trimmed plant Average 1.15 0.31 NANA NA NA NA NA NA NA weight (kg) Range (0-1.32) (0-1.1) Length of outerAverage 28.2 12.4 NA NA NA NA NA NA NA NA petioles to the Range 0-35(0-27) joint (cm) % Survival 81% 37% 16% 24% 31% 8% 9% 18% 17% 10% %Marketable 80% 35%  0%  0%  0%  0%  0%  0%  0%  0%

TABLE 5B Floribelle ADS-23 TBG 31 TBG 54 ADS-2 ADS-8 TBG 35 TBG 32 TBG53 Plant height (cm) Average NA NA NA NA NA NA NA NA NA Range Wholeplant Average NA NA NA NA NA NA NA NA NA weight (kg) Range Trimmed plantAverage NA NA NA NA NA NA NA NA NA weight (kg) Range Length of outerAverage NA NA NA NA NA NA NA NA NA petioles to the joint Range (cm) %Survival 21% 15% 4% 23% 42% 12% 33% 39% 52% % Marketable  0%  0% 0%  0% 0%  0%  0%  0%  0%

As shown in Tables 5A and 5B, under the fusarium conditions of thistrial most varieties had a few to several plants that were still aliveat evaluation but were not able to sustain development of marketablecelery stalks. Most were barely alive. However, TBG 45 and TBG 43 wereeach able to produce marketable celery stalks. TBG 43 was fairlytolerant (35% marketable) and essentially all survivors were marketable.Essentially all survivors of TBG 45 were marketable and at 80%marketable showed good tolerance for race 4.

Table 6 shows the results of a trial comparing characteristics of celerycultivar TBG 45 to celery varieties TBG 43, TBG 29, ADS-1, TBG 34, TBG37, TBG 28, TBG 33, ADS-20, and Command. The trial was transplanted inOxnard, California on Apr. 17, 2020, and evaluated on Jul. 7, 2020 (81days). This trial was grown in a production block that has beendeveloped with especially high levels of Fusarium oxysporum race 4 inorder to evaluate and develop varieties for increased tolerance to thedisease. Not only is this field especially high in the disease, but itis also the location where the disease was first identified. The plantpopulation (58,080 plants to the acre) was higher than the commercialnorm of approximately 45,000 to 47,000 plants to the acre. In order toincrease the impact of the disease in the spring, this trial wastransplanted late in the season when the soil temperature (activetemperature for the disease) was higher and the celery was not able toreach full maturity due to a mandated celery free window in VenturaCounty. Table 6, column 1 shows the characteristic and columns 2-11 showthe results for TBG 45, TBG 43, TBG 29, ADS-1, TBG 34, TBG 37, TBG 28,TBG 33, ADS-20, and Command, respectively. NA indicates data notavailable.

TABLE 6 TBG 45 TBG 43 TBG 29 ADS-1 TBG 34 TBG 37 TBG 28 TBG 33 ADS-20Command Plant height (cm) Average 57.4 23.20 NA NA NA 3.4 NA NA NA NARange (54-60) (0-49) (0-34) Whole plant Average 0.6 0.20 NA NA NA 0.0 NANA NA NA weight (kg) Range (0.46-0.7) (0-0.46) (0-0.14) Number of outerAverage 9.30 1.10 NA NA NA NA NA NA NA NA petioles Range (8-10) (0-5)(>35.6 cm) Number of inner Average 4.4 3.0 NA NA NA NA NA NA NA NApetioles Range (4-5) (0-7) (<35.6 cm) Length of outer Average 22.2 9.1NA NA NA 1.3 NA NA NA NA petioles to the Range (19.7-24) (0-20) (0-12.7)joint (cm) % Survival  95% 46% 5% 0% 0% 6% 0% 0% 0% 0% % Marketable 100%40% 0% 0% 0% 0% 0% 0% 0% 0%

As shown in Table 6, under the fusarium conditions of this trial, TBG 29and TBG 37 each had a few plants that survived, but only TBG 45 and TBG43 had marketable potential. TBG 43 was moderately tolerant (46%marketable) and TBG 49 had good tolerance (95% marketable). No otherconventional class celery showed tolerance.

Tables 7A, 7B, and 7C show the results of a trial comparingcharacteristics of celery cultivar TBG 45 to celery varieties TBG 43,TBG 29, ADS-1, TBG 34, TBG 37, TBG 28, TBG 33, ADS-22, Hill's Special,ADS-20, Challenger, Sonora, Conquistador, Command, Mission, Floribelle,ADS-23, TBG 31, TBG 54, TBG 35, TBG 32, TBG 53, Tall Utah 52-70 ‘R’Strain, and Tall Utah 52-75. The trial was transplanted in Oxnard,California on Aug. 19, 2020, and evaluated on Dec. 2, 2020 (105 days).This trial was grown in a production block that has been developed withespecially high levels of Fusarium oxysporum race 4 levels in order toevaluate and develop varieties for increased tolerance to the disease.Not only is this field especially high in the disease, but it is alsothe location where the disease was first identified. The plantpopulation (58,080 plants to the acre) was higher than the commercialnorm of approximately 45,000 to 47,000 plants to the acre. Table 7A,column 1 shows the characteristic and columns 2-10 show the results forTBG 45, TBG 43, TBG 29, ADS-1, TBG 34, TBG 37, TBG 28, TBG 33, andADS-22, respectively. Table 7B, column 1 shows the characteristic andcolumns 2-9 show the results for Hill's Special, ADS-20, Challenger,Sonora, Conquistador, Command, Mission, and Floribelle, respectively.Table 7C, column 1 shows the characteristic and columns 2-9 show theresults for ADS-23, TBG 31, TBG 54, TBG 35, TBG 32, TBG 53, Tall Utah52-70 ‘R’ Strain, and Tall Utah 52-75, respectively. NA indicates datanot available.

TABLE 7A TBG 45 TBG 43 TBG 29 ADS-1 TBG 34 TBG 37 TBG 28 TBG 33 ADS-22Plant height (cm) Average 70.3 40.5 8.8 5.5 1.9 0.6 0.0 0.0 3.7 Range(68-74) (0-70) (0-49) (0-25) (0-18) (0-9) (0-0) (0--0) (0-34) Wholeplant Average 0.87 0.37 0.06 NA NA NA NA NA NA weight (kg) Range(0.74-1.01) (0-0.82) (0-0.39) Trimmed plant Average 0.69 0.30 0.05 NA NANA NA NA NA weight Range (0.6-0.81) (0-0.67) (0-0.36) (kg @ 35.6 cm)Number of outer Average 9.9 4.4 0.7 NA NA NA NA NA NA petioles Range(8-11) (0-9) (0-4) (>35.6 cm) Number of inner Average 6.7 5.5 1.2 NA NANA NA NA NA petioles Range (6-8) (0-10) (0-6) (<35.6 cm) Length of outerAverage 28.8 14.5 2.7 3.2 0.7 0.1 0.0 0.0 1.1 petioles to the Range(26-32) (0-27) (0-13.7) (0-12) (0-7) 90-2) (0-0) (0-0) (0-11) joint (cm)Width of outer Average 22.9 10.7 3.1 NA NA NA NA NA NA petioles at theRange (20.3-25) (0-20) (0-16) midrib (mm) Thickness of outer Average10.3 5.7 1.7 NA NA NA NA NA NA petioles at the Range (9.3-11.3) (0-11.3)(0-9) midrib (mm) Petiole smoothness Smooth/ Smooth/ Slight rib NA NA NANA NA NA Slight rib Slight rib Petiole cup Cup Cup/Deep Cup NA NA NA NANA NA cup % Marketable 94% 50%  0%  0%  0%  0%   0%   0%  0% Disease:Survival rate (%) Healthy 95% 74% 18%  0%  0%  0%   0%   0%  2% Dying 3%  3% 10% 17% 12%  7%   0%   0% 13% Dead  3% 23% 82% 83% 88% 93% 100%100% 85% Overall fusarium Average 4.5 2.5 1.5 0 0 0 0 0 0 ratings (0 =dead, Range 4-5 (0-5) (0-5) 0-1 (0-1) 0-1 (0-0) (0-0) (0-3) 5 =tolerant)

TABLE 7B Hill′s Special ADS-20 Challenger Sonora Conquistador CommandMission Floribelle Plant height (cm) Average 1.2 1.5 NA NA NA NA NA 0.0Range (0-18) (0-21) (0-0) Whole plant weight Average NA NA NA NA NA NANA NA (kg) Range Trimmed plant Average NA NA NA NA NA NA NA NA weightRange (kg @ 35.6 cm) Number of outer Average NA NA NA NA NA NA NA NApetioles (>35.6 cm) Range Number of inner Average NA NA NA NA NA NA NANA petioles Range (<35.6 cm) Length of outer Average 0.6 0.5 NA NA NA NANA 0.0 petioles to the joint Range (0-8) (0-7) (0-0) (cm) Width of outerAverage NA NA NA NA NA NA NA NA petioles at the Range midrib (mm)Thickness of outer Average NA NA NA NA NA NA NA NA petioles at the Rangemidrib (mm) Petiole smoothness NA NA NA NA NA NA NA NA Petiole cup NA NANA NA NA NA NA NA % Marketable  0%  0%  0%  0%  0%  0%  0%   0% Disease:Survival rate (%) Healthy  0%  0%  0%  0%  0%  0%  0%   0% Dying 13% 10%13% 11%  8% 15% 10%   0% Dead 87% 90% 87% 89% 92% 85% 90% 100% Overallfusarium Average 0 1 0 0 0 0 0 0 ratings (0 = dead, Range 0-1 (0-1) 0-10-1 0-1 0-1 0-1 (0-0) 5 = tolerant)

TABLE 7C Tall Utah 52-70 ′R′ Tall Utah ADS-23 TBG 31 TBG 54 TBG 35 TBG32 TBG 53 Strain 72-75 Plant height (cm) Average 0.7 2.9 0.0 0.0 0.5 0.5NA NA Range (0-18) (0-28) (0-0) (0-0) (0-15) (0-11) Whole plant weightAverage NA NA NA NA NA NA NA NA (kg) Range Trimmed plant Average NA NANA NA NA NA NA NA weight Range (kg @ 35.6 cm) Number of outer Average NANA NA NA NA NA NA NA petioles (>35.6 cm) Range Number of inner AverageNA NA NA NA NA NA NA NA petioles Range (<35.6 cm) Length of outerAverage 0.3 1.1 0.0 0.0 0.2 0.1 NA NA petioles to the joint Range (0-5)(0-12) (0-0) (0-0) (0-5) (0-3) (cm) Width of outer Average NA NA NA NANA NA NA NA petioles at the Range midrib (mm) Thickness of outer AverageNA NA NA NA NA NA NA NA petioles at the Range midrib (mm) Petiolesmoothness NA NA NA NA NA NA NA NA Petiole cup NA NA NA NA NA NA NA NA %Marketable  0%  0%   0%   0%  0%  0%  0%  % Disease: Survival rate (%)Healthy  0%  2%   0%   0%  0%  0%  0%  0% Dying  4% 13%   0%   0%  5% 5%  7%  1% Dead 96% 85% 100% 100% 95% 95% 93% 99% Overall fusariumAverage 0 0 0 0 0 0 0 0 ratings (0 = dead, Range 0-1 (0-2) (0-0) (0-0)(0-1) (0-1) 0-1 0-1 5 = tolerant)

As shown in Tables 7A, 7B, and 7C, under the fusarium conditions of thistrial, TBG 29 had a few plants that had slightly better vigor, but withonly 18% survival and an average fusarium rating of 1.5 even thestrongest of the plants were not marketable. Under the fusariumconditions of this trial which has been developed to be stronger thantraditionally infected fields, the tolerance of TBG 43 is starting tobreak down with 74% survival and 50% marketable. TBG 45, which wasspecifically bred for tolerance to race 4, had 94% marketability and wasvery tolerant (4-5 rating). No other variety showed any potential underthese conditions.

Tables 8-9 show the results for trials that were produced in springconditions in Oxnard, California when there were either low levels offusarium race 2 or slight bolting pressure or both present concurrently.With spring weather conditions able to impact varieties for either orboth conditions, the varieties tolerances can have a significant impacton marketability of the product and success for the producer.

Under these conditions, celery cultivar TBG 45 showed no impact fromfusarium or bolting, usually held well in the top tier of varieties foryield based on trimmed plant weight and was generally most similar toTBG 43 and TBG 29. However, TBG 45 did have a higher incidence ofsuckers (20%) in both trials compared to TBG 43, and TBG 45 and was moresimilar to TBG 29 for risk of feather leaf (Table 9). In one trial TBG45 outperformed all varieties for yield as measured by trimmed plantweight (Table 8) and in the second it was very similar to TBG 43 andlower than TBG 29 (Table 9).

Tables 8A, 8B, and 8C show the results of a trial comparingcharacteristics of celery cultivar TBG 45 to celery varieties TBG 43,TBG 29, ADS-1, TBG 34, TBG 37, TBG 28, TBG 33, Hill's Special, ADS-20,ADS-22, Challenger, Sonora, Conquistador, Command, Mission, ADS-23, TBG31, TBG 54, ADS-8, TBG 35, TBG 32, TBG 53, Tall Utah 52-70 ‘R’ Strain,and Tall Utah 52-75. The trial was transplanted in Oxnard, California onMar. 21, 2020, and evaluated on Jun. 28, 2020 (99 days) in a normalproduction field with moderate pressure due to Fusarium oxysporum race 2and slight bolting pressure. Tall Utah 52-70 ‘R’ Strain, considered thesusceptible check for both traits, was significantly impacted byfusarium while slightly impacted by bolting. The plant population(58,080 plants to the acre) was higher than the commercial norm ofapproximately 45,000 to 47,000 plants to the acre. Table 8A, column 1shows the characteristic and columns 2-10 show the results for TBG 45,TBG 43, TBG 29, ADS-1, TBG 34, TBG 37, TBG 28, TBG 33, and Hill'sSpecial, respectively. Table 8B, column 1 shows the characteristic andcolumns 2-9 show the results for ADS-20, ADS-22, Challenger, Sonora,Conquistador, Command, Mission, and ADS-23, respectively. Table 8C,column 1 shows the characteristic and columns 2-9 show the results forTBG 31, TBG 54, ADS-8, TBG 35, TBG 32, TBG 53, Tall Utah 52-70 ‘R’Strain, and Tall Utah 52-75, respectively. NA indicates data notavailable.

TABLE 8A Hill′s TBG 45 TBG 43 TBG 29 ADS-1 TBG 34 TBG 37 TBG 28 TBG 33Special Plant height Average 82.7 88.6 82.9 76.9 76.50 78.80 96.00 78.2065.50 (cm) Range (81-85) (82-94) (80-89) (70-81) (72-82) (76-82)(92-100) (68-85) (56-70) Whole plant Average 1.66 1.49 1.51 1.23 1.181.25 1.27 1.18 1.02 weight (kg) Range (1.3-2.12) (1.24-1.76) (1.12-2.14)(0.68-1.72) (1-1.34) (1.02-1.5) (1.1-1.5) (0.84-1.44) (0.66-1.36)Trimmed plant Average 1.3 1.09 1.17 0.60 0.87 0.94 1.01 0.53 0.79 weight(kg) Range (1.02-1.7) (0.9-1.3) (0.9-1.56) (0-1.32) (0-1.12) (0.74-1.06)(0-1.5) (0-1.16) (0-1.8) Number of Average 12.0 10.4 11.0 6.2 7.4 11.610.6 5 7.8 outer petioles Range (9-14) (8-13) (10-12) (0-13) (0-10)(10-13) (0-16) (0-12) (0-12) Number of Average 5.2 4.8 5.3 3.1 4.2 5.15.4 2.5 4 inner petioles Range (3-6) (4-6) (4-6) (0-6) (0-5) (4-6) (0-8)(0-6) (0-6) Length of Average 32.3 28.1 28.6 24.5 28.6 28.4 33.3 28.025.4 outer Range (28-36) (26.7-29) (27-31) (22-27.3) (27-30) (26.3-33)(30-37) (23-33) (22.3-27.7) petioles to the joint (cm) Width of outerAverage 24.9 26.1 25.4 14.6 22.9 22.4 20.9 13.2 19.3 petioles at theRange (22.3-28.3) (25-28.3) (24-28.7) (0-26) (0-27) (21-23.7) (0-27)(0-27.7) (0-28.3) midrib (mm) Thickness of Average 12.2 12.7 12.7 6.910.6 10.5 9.9 6.3 8.7 outer petioles Range (11.3-12.7) (11.3-13.7)(11.7-14) (0-12.7) (0-13) (7-12.3) (0-13.3) (0-13.3) (0-11.3) at themidrib (mm) Length of Average 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 seedstems Range (0-0) (0-0) (0-0) (0-0) (0-0) (0-0) (0-0) (0-0) (0-0) (cm)Median 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Petiole color 5 gy 6/6 5 gy6/8, 5 gy 6/6 5 gy 6/6 5 gy 6/6 5 gy 6/4 5 gy 6/6 5 gy 6/6 5 gy 7/4,(Munsell color) 5 gy 3/4 5 gy 6/4 Leaf color 5 gy 3/4 5 gy 3/4 5 gy 3/45 gy 3/4 5 gy 3/4 5 gy 3/4 5 gy 3/4 5 gy 3/4 5 gy 3/4 (Munsell color)Petiole smoothness Slight rib Smooth Smooth/ Smooth Slight Smooth SmoothSmooth/ Smooth Slight rib rib/Rib Slight rib Petiole cup Cup Cup Cup CupCup Cup Cup Cup Cup % Marketable 100% 100% 100% 60% 100% 100% 80% 50% 80% Disease ratings: Overall Average 5 5 5 3.5 5 5 5 4 3.5 fusariumRange 5 5 5 3-5 5 5 5 3-5 3-4 ratings (0 = dead, 5 = tolerant) RootAverage 5 5 4 3 3.5 4 3.5 2.5 2.5 fusarium Range 4-5 4-5 4-5 1-4 2-4 42-5 1-4 2-3 ratings (0 = dead, 5 = tolerant) Defects: % Node crack   0%  0%   0%  0%   0%   0%  0% 30% 100% % Brown stem   0%   0%   0%  0%  0%   0%  0%  0%   0% % Butt crack   0%   0%   0% 10%   0%   0% 10% 30% 20% % Top pith   0%   0%   0% 10%  10%   0%  0% 50%   0% % Butt pith  0%  70%   0% 80% 100%  20%  0% 50%  50% % Suckers  20%   0%   0%  0%  0%   0%  0% 20% 100% % Feather leaf   0%   0%   0%  0%   0%   0%  0%40%   0% % Twist   0%  20%   0%  0%  40%   0% 10% 30% 100%

TABLE 8B ADS-20 ADS-22 Challenger Sonora Conquistador Command MissionADS-23 Plant height (cm) Average 67.60 84.6 74.2 67.1 64.3 71.80 70.751.40 Range (61-73) (81-89) (37-90) (62-72) (23-74) (63-77) (65-75)(32-70) Whole plant Average 1.00 1.35 1.07 1.03 0.80 1.20 0.98 0.43weight (kg) Range (0.6-1.28) (0.98-1.52) (0.08-1.52) (0.46-1.44)(0.06-1.3) (0.92-1.42) (0.68-1.26) (0.1-0.86) Trimmed plant Average 0.671.08 0.50 0.45 0.60 1.04 0.26 NA weight (kg) Range (0-1.1) (0.8-1.18)(0-1.22) (0-1.22) (0-1.56) (0.8-1.28) (0-1.06) Number of outer Average6.5 10.9 4.8 4.9 5.1 10.0 2.8 NA petioles Range (0-11) (9-13) (0-11)(0-14) (0-13) (9-11) (0-10) Number of inner Average 3.5 5.5 2.8 4.4 3.26.7 1.7 NA petioles Range (0-6) (4-7) (0-7) (0-14) (0-8) (6-8) (0-7)Length of outer Average 26.2 24.4 29.0 25.1 23.9 25.7 27.8 16.4 petiolesto the Range (21.3-30.3) (22.7-25.7) (16-34.7) (18.3-27) (8-29)(21.3-29.7) (26-30.7) (10.3-25.3) joint (cm) Width of outer Average 17.225.2 11.2 8.4 11.3 24.7 6.8 NA petioles at the Range (0-26) (23-27.3)(0-26) (0-22.3) (0-23.7) (22-27.7) (0-21) midrib (mm) Thickness ofAverage 8.1 12.0 5.6 3.9 5.3 11.8 2.8 NA outer petioles at Range(0-12.7) (10.7-13) (0-12.7) (0-10.3) (0-11.3) (10-14.7) (0-10) themidrib (mm) Length of seed Average 0.0 0.0 0.0 0.0 0.0 0.0 0.0 2.3 stems(cm) Range (0-0) (0-0) (0-0) (0-0) (0-0) (0-0) (0-0) (0-23) Median 0.00.0 0.0 0.0 0.0 0.0 0.0 0.0 Petiole color 5 gy 7/4, 5 gy 6/6 5 gy 6/6 5gy 6/6 5 gy 6/6 5 gy 6/6 5 gy 6/6 NA (Munsell color) 5 gy 6/4 Leaf color5 gy 3/4 5 gy 3/4 5 gy 3/4 5 gy 4/4 5 gy 4/4 5 gy 4/4 5 gy 4/4 NA(Munsell color) Petiole smoothness Smooth Smooth Smooth/ Slight ribSmooth Slight rib Slight rib NA Slight rib Petiole cup Cup Cup Cup CupCup Cup Cup NA % Marketable  70% 100%  40%  40%  50% 100%  30%   0%Disease ratings: Overall fusarium Average 4 5 4.5 3.5 3.5 5 3.5 1ratings (0 = dead, Range 3-4 5 1-5 2-5 1-5 5 2-5 1-2 5 = tolerant) Rootfusarium Average 2.5 4 3 2.5 2.5 2.5 1.5 1 ratings (0 = dead, Range 1-33-4 1-4 1-4 1-4 1-4 1-4 1 5 = tolerant) Defects: % Node crack 100%  10%  0% 10%  0%  50%  0%   0% % Brown stem   0%   0%  40% 30%  0%   0% 40% 90% % Butt crack  30%  10%  50% 40% 30%   0% 30% 100% % Top pith  30%  0%  50% 60% 50%  40% 70% 100% % Butt pith  50%  50%   0% 90% 90%  50%90% 100% % Suckers  80%   0%  10%  0% 30%   0%  0% 100% % Feather leaf  0%   0%  30% 60% 70%  20% 40%  70% % Twist 100%  40% 100% 60% 50%  20%50% 100%

TABLE 8C Tall Utah 52-70 Tall ′R′ Utah TBG 31 TBG 54 ADS-8 TBG 35 TBG 32TBG 53 Strain 52-75 Plant height (cm) Average 69.40 64.00 52.80 49.7069.90 75.80 54.8 65.8 Range (32-83) (59-70) (36-77) (28-92) (63-79)(72-79) (46-62) (57-72) Whole plant Average 1.28 0.93 0.34 0.45 1.031.34 0.68 0.82 weight (kg) Range (0.14-1.94) (0.7-1.26) (0.1-0.96)(0.12-1.08) (0.48-1.94) (1-1.68) (0.2-1.48) (0-1.24) Trimmed plantAverage 0.50 0.28 NA NA 0.17 0.85 NA 0.44 weight (kg) Range (0-1.44)(0-1.06) (0-1) (0-1.32) (0-1.04) Number of outer Average 4.7 2.7 NA NA2.5 10.2 NA 4.4 petioles Range (0-13) (0-10) (0-14) (0-15) (0-10) Numberof inner Average 2.2 1.3 NA NA 0.9 4.1 NA 2.7 petioles Range (0-6) (0-5)(0-6) (0-7) (0-6) Length of outer Average 27.9 29.2 13.9 15.3 29.3 32.120.5 25.0 petioles to the Range (11.7-36) (22-66) (10.7-20) (9.3-22.3)(16.7-35.7) (28.7-36.7) (16-24) (19.3-29.3) joint (cm) Width of outerAverage 10.0 7.1 NA NA 3.9 18.0 NA 11.2 petioles at the Range (0-26.3)(0-25) (0-19.3) (0-24) (0-24) midrib (mm) Thickness of Average 5.0 3.5NA NA 2.1 9.0 NA 5.4 outer petioles at Range (0-13) (0-12.7) (0-11.3)(0-12.7) (0-12) the midrib (mm) Length of seed Average 18.4 0.0 29.312.7 20.2 0.0 6.4 0.0 stems (cm) Range (0-83) (0-0) (0-77) (0-92) (0-64)(0-0) (0-30) (0-0) Median 0.0 0.0 15.5 2.0 12.5 0.0 1.5 0.0 Petiolecolor 5 gy 7/4- 5 gy 7/4- NA NA NA 5 gy 7/4- NA 5 gy 6/6 (Munsell color)5 gy 6/6 5 gy 6/6 5 gy 6/6 Leaf color 5 gy 3/4 5 gy 3/4 NA NA NA 5 gy3/4 NA 5 gy 4/4 (Munsell color) Petiole smoothness Slight Smooth/ NA NASmooth Slight NA Smooth rib/Rib Slight rib rib/Rib Petiole cup Cup CupNA NA Slight cup Slight NA Cup cup/Cup % Marketable 20%  30%   0%   0%10%  80%   0% 50% Disease ratings: Overall fusarium Average 4 3.5 1 13.5 4 1.5 3 ratings (0 = dead, Range 1-5 3-4 0-2 1-2 3-4 4-5 0-3 1-5 5 =tolerant) Root fusarium Average 2.5 2 1 1 2 3 1.5 2 ratings (0 = dead,Range 1-3 1-3 0-1 1-2 1-3 3 0-3 1-3 5 = tolerant) Defects: % Node crack 0%   0%   0%   0%  0%   0%   0%  0% % Brown stem 30%   0%  80%  40%  0%  0%  60%  0% % Butt crack 60%  30% 100% 100% 50%   0% 100% 40% % Toppith 60%  70% 100% 100% 90%   0% 100% 70% % Butt pith 70% 100% 100% 100%90% 100% 100% 90% % Suckers 30%   0% 100% 100% 60%   0% 100%  0% %Feather leaf 50%  70%   0%   0% 40%  30% 100% 50% % Twist 60%   0% 100%100% 60%   0% 100% 50%

As shown in Tables 8A, 8B, and 8C, celery cultivars TBG 45 and TBG 43were the most tolerant to Fusarium oxysporum race 2 based on both theoverall and root infection ratings, in addition to the percentmarketability followed closely by TBG 29 and TBG 37, and slightly moredistantly TBG 28. Of these, TBG 28 had was the tallest for plant heightfollowed by TBG 45, and the others were fairly similar. TBG 45 and TBG28 were very similar with longer petiole length compared to TBG 43, TBG29, and TBG 37. Except for Tall Utah 52-70 ‘R’ Strain, bolting pressurewas not notable in what most would consider the California coastalvarieties. However, it was a significant issue in varieties developedfor Florida production including ADS-8, TBG 31, TBG 32, and ADS-23. Itwas not observed in two Florida varieties TBG 54 and TBG 53.

Table 9 shows the results of a trial comparing characteristics of celerycultivar TBG 45 to celery varieties TBG 43, TBG 29, ADS-1, TBG 34, TBG37, TBG 28, TBG 31, TBG 54, TBG 35, TBG 32, and TBG 53. The trial wastransplanted in Oxnard, California on Mar. 8, 2021, and evaluated onJun. 25, 2021 (109 days) in a normal production field with very lowpressure due to Fusarium oxysporum race 2, but slight pressure forbolting (442 hours below 50° F.). The plant population (58,080 plants tothe acre) was higher than the commercial norm of approximately 45,000 to47,000 plants to the acre. Table 9, column 1 shows the characteristicand columns 2-13 show the results for TBG 45, TBG 43, TBG 29, ADS-1, TBG34, TBG 37, TBG 28, TBG 31, TBG 54, TBG 35, TBG 32, and TBG 53,respectively. NA indicates data not available.

TABLE 9 TBG TBG TBG ADS- TBG TBG TBG TBG TBG TBG TBG TBG 45 43 29 1 3437 28 31 54 35 32 53 Plant height Average 83.4 88.9 80.5 80.2 80.4 89.097.5 43.6 48.8 NA 35.4 91.8 (cm) Range (82-88) (86-92) (78-83) (75-85)(77-84) (83-95) (94-104) (0-89) (0-85) (0-91) (89-95) Whole plantAverage 1.76 1.77 1.87 1.69 1.27 1.53 1.72 0.85 0.77 NA 0.59 1.20 weight(kg) Range (1.46- (1.48- (1.6- (1.46- (0.98- (1.18- (1.32- (0-2) (0-(0-1.72) (0-1.7) 2.1) 2.02) 2.28) 2.12 1.62) 1.86) 2.12) 1.55) Trimmedplant Average 1.29 1.27 1.42 1.31 0.94 1.14 1.25 0.48 0.57 NA 0.43 0.81(kg @ 35.6 cm) Range (1.06- (1.1- (1.22- (1.1- (0.76- (0.82- (0.94- (0-(0- (0-1.24) (0- weight 1.52) 1.46) 1.74) 1.6) 1.18) 1.44) 1.5) 1.38)1.18) 1.16) Number of Average 12.1 11.2 12.0 10.5 9.6 11.7 12.9 4.9 6.8NA 4.4 9.5 outer petioles Range (10-14) (10-12) (10-14) (9-12) (8-11)(9-13) (11-16) (0-14) (0-13) (0-14) (0-13) (>35.6 cm) Number of Average4.5 5.2 4.9 6.1 3.3 5.2 5.6 1.7 2.2 NA 4.4 3.3 inner petioles Range(3-5) (4-6) (4-8) (5-7) (3-4) (4-6) (4-7) (0-6) (0-5) (0-14) (0-5)(<35.6 cm) Length of outer Average 31.3 30.2 26.9 29.4 31.0 34.4 38.913.7 17.9 NA 14.0 27.5 petioles to the Range (28- (28- (18- (26.3-(28.7- (30.7- (36.3- (0- (0- (0-37) (0-38) joint (cm) 36.3) 31.7) 30.3)31.7) 33.7) 38.7) 40.7) 37.3) 33.3) Width of outer Average 27.3 28.330.3 26.8 27.1 25.3 28.1 10.1 14.2 NA 10.6 19.7 petioles at the Range(24.3- (25.3- (28.7- (22.3- (26- (22.3- (26.7- (0- (0- (0-29.7) (0-midrib (mm) 29) 30.3) 33.7) 29.3) 28.7) 27) 29.7) 26.7) 25.7) 25.7)Thickness of Average 11.0 12.5 12.6 12.0 11.0 10.3 11.9 4.7 6.5 NA 4.78.6 outer petioles Range (10-12) (12- (11.3- (10.3- (9.7- (9.3- (11-13)(0- (0- (0-12.3) (0-12) at the midrib 13.3) 14) 13) 12.3) 11.7) 13.3)12.7) (mm) Seed stem Average 0.0 0.0 0.0 1.3 0.0 0.5 0.0 43.7 23.2 118.768.3 13.1 length (cm) Range (0-0) (0-0) (0-0) (0-11) (0-0) (0-4) (0-0)(0-96) (0-71) (99-140) (0-131) (0-80) Median 0.0 0.0 0.0 0.0 0.0 0.0 0.053.5 16.0 118.0 84.0 0.8 Petiole color 5 gy 5 gy 5 gy 5 gy 5 gy 5 gy 5gy 5 gy 5 gy NA 5 gy 5 gy (Munsell color) 6/6 6/6 7/6 6/6 6/6 6/6 6/67/6 7/6 7/6 6/6 Leaf color 5 gy 5 gy 5 gy 5 gy 5 gy 5 gy 5 gy 5 gy 5 gyNA 5 gy 5 gy (Munsell color) 3/4 3/4 3/4 3/4 3/4 3/4 3/4 4/4 4/4 4/4 4/4Petiole smoothness Slight Smooth Smooth/ Smooth Smooth Smooth SmoothSmooth Smooth NA Smooth Smooth rib/ Slight Rib rib Petiole cup SlightCup Slight Cup/ Cup Cup Cup Cup TBG NA Slight Slight cup/ cup Deep 54Cup cup cup Cup cup % Marketable 100% 100% 100% 100% 100% 100% 100%   0%  0% 0%  30% 80% Defects: % Top pith   0%   0%   0%   0% 100%   0%   0%100% 100% NA 100% 20% % Butt pith   0%   0%   0%   0%   0%   0%   0%100%  40% NA  80% 20% % Node crack   0%   0%  20%   0%  30%   0%   0%  0%   0% NA   0% 10% % Butt crack   0%   0%   0%   0%   0%   0%   0% 10%   0% NA   0%  0% % Suckers  20%   0%   0%   0%  10%   0%   0%  40% 40% NA  60% 20% % Feather leaf  20%   0%  30%   0%   0%   0%   0%   0%  0% NA   0%  0% % Twist   0%  10%   0%   0%  20%  40% 100%  40%   0% NA 60% 20% % Black heart   0%   0%   0%   0%   0%   0%   0%  10%   0% NA  0%  0% % Brown stem   0%   0%   0%   0%   0%  10%   0%  30%  80% NA 70% 20%

As shown in Table 9, under the slight bolting pressure that this trialexperienced, varieties specially developed for production in Florida hadsignificant seed stem development (TBG 35, TBG 31, TBG 32, TBG 54).Except for ADS-1, there was very little impact on the remainingvarieties which had been developed for production in California. Underthese conditions, TBG 29 out yielded all varieties (trimmed plantweight), and TBG 45, TBG 43, ADS-1, and TBG 28 were very similar andnext in line for trimmed weight. However, TBG 29 and TBG 45 wereimpacted more by feather leaves. TBG 28 performed as it would normally,producing the tallest plants with the longest petioles to the jointfollowed by TBG 37; however, both also had more petiole twisting. TBG45, TBG 29, and TBG 28 were similar in producing an average of 12 ormore petioles longer than 35.6 cm. ADS-1 and TBG 43 appeared to be mostsimilar with one another for most of their characteristics and the twothat were most free from defects. NA indicates that the data for thatparticular item was not available, in this case for TBG 35 due to severeseed stem development and distortion.

Table 10 shows the results for a trial grown in Ventura County duringwhat is prominently referred to as the bolting window; the period oftime when conditions frequently cause initiation of bolting and thedegree of bolting susceptibility may be measured by the seed stemlength, allowing for comparison of different varieties. This windowtypically includes the coldest and shortest day length period of theyear and usually comprises the months of December, January, and Februaryin the earlier part of the cropping cycle. This trial shows the resultsfor celery grown with 1,061 hours at or below 50° F. While this numberof hours is traditionally fairly high and associated with severebolting, the level of bolting in this trial was actually fairlymoderate. This could be due to the distribution of the cold hours andpotential periods of warm weather which could have a negative impact onthe seed stem development. Cold weather at the last stages of productioncan often slow seed stem development within the plant just prior tomarket stage.

Under these conditions, celery cultivar TBG 45 was very similar to TBG43 for slight bolting tolerance and not quite as good as TBG 29 which isconsidered to have more moderate tolerance. TBG 45 cannot compare toADS-20 and TBG 34, both which are considered very tolerant. Suckersremain a defect concern for TBG 45 in the spring when there is somepressure for bolting.

Tables 10A and 10B show shows the results of a trial comparingcharacteristics of celery cultivar TBG 45 to celery varieties TBG 43,TBG 29, ADS-1, TBG 34, TBG 37, TBG 28, TBG 33, Hill's Special, ADS-20,Challenger, Sonora, Conquistador, Mission, Command, Floribelle, ADS-23,TBG 31, TBG 54, ADS-8, TBG 35, TBG 32, and TBG 53. The trial wastransplanted in Oxnard, California on Dec. 19, 2020, and evaluated onApr. 27, 2021 (129 days). This trial was grown in a normal productionfield that had moderate to heavy pressure for bolting (1,061 hours below50° F.), measured by seed stem length. The only characteristics measuredfor varieties with an average seed stem length of 10 cm and higher werelength of outer petioles at the joint and seed stem length. The plantpopulation (58,080 plants to the acre) was higher than the commercialnorm of approximately 45,000 to 47,000 plants to the acre. Table 10A,column 1 shows the characteristic and columns 2-13 show the results forTBG 45, TBG 43, TBG 29, ADS-1, TBG 34, TBG 37, TBG 28, TBG 33, Hill'sSpecial, ADS-20, Challenger, and Sonora, respectively. Table 10B, column1 shows the characteristic and columns 2-12 show the results forConquistador, Mission, Command, Floribelle, ADS-23, TBG 31, TBG 54,ADS-8, TBG 35, TBG 32, and TBG 53, respectively. NA indicates data notavailable.

TABLE 10A TBG TBG TBG ADS- TBG TBG TBG TBG Hill′s ADS- 45 43 29 1 34 3728 33 Special 20 Challenger Sonora Plant height Average 81.7 84.6 1.65NA 76.2 83.8 NA NA 71.5 73.0 NA NA (cm) Range (76-85) (68-93)(1.16-2.04) (72-80) (77-91) (69-76) (70-76) Whole plant Average 1.551.57 1.65 NA 1.57 1.46 NA NA 1.25 1.21 NA NA weight (kg) Range (1.02-2)(1.282)- (1.16-2.04) (1.06- (0.96- (1.08- (1-1.58) 1.84) 2.06) 1.56)Trimmed Average 1.17 1.17 1.31 NA 1.13 1.23 NA NA 1.08 0.94 NA NA plantweight Range (0.78- (0.96- (0.96- (0.74- (0.84- (0.96- (0.74- (kg) 1.48)1.5) 1.62) 1.36) 1.64) 1.34) 1.22) Number of Average 12.2 11.2 14.2 NA11.5 NA NA NA 9.2 11.6 NA NA outer petioles Range (11-14) (10-13)(13-15) (9-13) (8-12) (10-13) Number of Average 8.4 9.3 10.4 NA 9.2 NANA NA 9.2 8.7 NA NA inner petioles Range (6-12) (8-10) (9-13) (7-12)(8-12) (7-10) Length of Average 43.1 36.0 35.0 33.3 33.1 39.2 44.1 44.334.6 34.3 41.8 36.8 outer petioles Range (36.7- (31.7- (31.3- (29.3-(29.7- (34-49) (39-50) (41- (31.3 (32.3- (37-47.7) (34.7- to the joint48) 38.7) 37.3) 37) 37) 47.3) -37.7) 36.7) 38.7) (cm) Width of outerAverage 28.4 29.0 31.8 NA 28.7 NA NA NA 26.0 27.0 NA NA petioles at theRange (24.3- (25.3- (28.0- (26.3- (23.7- (22.7- midrib (mm) 32) 31.7)36.7) 32.3) 30.7) 29.3) Thickness of Average 9.9 11.3 10.6 NA 10.8 NA NANA 9.4 9.6 NA NA outer petioles Range (8.7- (9.7- (9.3- (9.7-12) (8.3-(8.3-11) at the midrib 11) 12.7) 11.3) 10.7) (mm) Seed stem Average 4.74.7 2.5 11.1 0 10.1 13.9 16.6 0.7 0 21.9 17.5 length (cm) Range (0-14)(0-16) (0-5) (4-21) (0-0) (0-22) (2-35) (8-24) (0-5) (0-0) (15-34)(6-35) Median 2.5 3.5 2.5 1 0 11.5 13.5 17.5 0 0 21 15 Petiole color 5gy 5 gy 5 gy NA 5 gy 5 gy NA NA 5 gy 5 gy NA NA (Munsell color) 6/6 6/46/6 6/6 6/4 6/6 6/6 Leaf color 5 gy 5 gy 5 gy NA 5 gy 5 gy NA NA 5 gy 5gy NA NA (Munsell color) 3/4 3/4 4/4 4/4 3/4 4/4 4/4 Petiole smoothnessSmooth/ Smooth Smooth/ NA Slight Slight NA NA Smooth Smooth NA NA RibSlight rib rib/Rib rib/Rib Petiole cup Cup Cup Slight NA Cup Slight NANA Cup Cup NA NA Cup cup/ Cup Defects: % Node crack  0% 0 0% NA  0%  30%NA NA  0% 0 NA NA % Butt crack  0%  0% 0% NA  0%  10% NA NA  0%  0% NANA % Suckers 70%  0% 0% NA 60% 100% NA NA  0% 20% NA NA % Twist 40% 30%0% NA 60% 100% NA NA 50%  0% NA NA % Feather leaf  0%  0% 0% NA  0%   0%NA NA  0%  0% NA NA

TABLE 10B Conquistador Mission Command Floribelle ADS-23 TBG 31 TBG 54ADS-8 TBG 35 TBG 32 TBG 53 Plant height Average NA NA NA NA NA NA NA NANA NA NA (cm) Range Whole plant Average NA NA NA NA NA NA NA NA NA NA NAweight (kg) Range Trimmed plant Average NA NA NA NA NA NA NA NA NA NA NAweight (kg) Range Number of Average NA NA NA NA NA NA NA NA NA NA NAouter petioles Range Number of Average NA NA NA NA NA NA NA NA NA NA NAinner petioles Range Length of outer Average 35.5 35.8 34.4 32.6 38.740.3 35.8 33.0 37.4 41.6 45.0 petioles to the Range (33.3-40) (33-39)(31-38.7) (28.7- (34.3- (37- (33.3- (28.3- (32.7- (38- (43.7- joint (cm)35.7) 43) 44.7) 42) 36.7) 41.3) 44.7) 46.7) Width of outer Average NA NANA NA NA NA NA NA NA NA NA petioles at the Range midrib (mm) Thicknessof Average NA NA NA NA NA NA NA NA NA NA NA outer petioles Range at themidrib (mm) Seed stem Average 16.7 10.3 16.6 56.1 11.9 38.7 24.6 42.660.4 40.1 37.6 length (cm) Range (8-33) (2-21) (10-31) (40-86) (0-21)(23-65) (15-40) (34-52) (38-83) (25-61) (23-49) Median 16 9.5 15 50.5 1637.5 23.5 42 60 39.5 37 Petiole color NA NA NA NA NA NA NA NA NA NA NA(Munsell color) Leaf color NA NA NA NA NA NA NA NA NA NA NA (Munsellcolor) Petiole smoothness NA NA NA NA NA NA NA NA NA NA NA Petiole cupNA NA NA NA NA NA NA NA NA NA NA % Node crack NA NA NA NA NA NA NA NA NANA NA % Butt crack NA NA NA NA NA NA NA NA NA NA NA % Suckers NA NA NANA NA NA NA NA NA NA NA % Twist NA NA NA NA NA NA NA NA NA NA NA %Feather leaf NA NA NA NA NA NA NA NA NA NA NA

As shown in Tables 10A and 10B, with the cold hours being so high onewould have expected considerably more seed stem development; however, itremained a little lower than the hours would suggest. There are manyenvironmental issues that can alter expected results including coldperiods being interrupted by warmer periods which may break the boltingtriggers. Full characteristic notes were only taken for more boltingtolerant cultivars (seed stem development of less than 10 cm). ADS-20and TBG 34 were the most bolting tolerant with no seed stem developmentunder these conditions followed by Hill's Special. TBG 29 was the nextmost bolting tolerant followed by TBG 45 and TBG 43 which were both verysimilar. TBG 29 had the highest yield based on average stalk weight,petiole count (outer) and the widest petioles. Next best yielding wereTBG 43 and TBG 45 based on weight which were similar to TBG 34; however,due to the expanded seed stem weight was likely misstated sinceconsiderable weight may be attributed to the seed stem compared to TBG34 which has none. TBG 45 was worse than TBG 43 for defects.

The use of the terms “a,” “an,” and “the,” and similar referents in thecontext of describing the invention (especially in the context of thefollowing claims) are to be construed to cover both the singular and theplural, unless otherwise indicated herein or clearly contradicted bycontext. The terms “comprising,” “having,” “including,” and “containing”are to be construed as open-ended terms (i.e., meaning “including, butnot limited to,”) unless otherwise noted. Recitation of ranges of valuesherein are merely intended to serve as a shorthand method of referringindividually to each separate value falling within the range, unlessotherwise indicated herein, and each separate value is incorporated intothe specification as if it were individually recited herein. Forexample, if the range 10-15 is disclosed, then 11, 12, 13, and 14 arealso disclosed. All methods described herein can be performed in anysuitable order unless otherwise indicated herein or otherwise clearlycontradicted by context. The use of any and all examples, or exemplarylanguage (e.g., “such as”) provided herein, is intended merely to betterilluminate the invention and does not pose a limitation on the scope ofthe invention unless otherwise claimed. No language in the specificationshould be construed as indicating any non-claimed element as essentialto the practice of the invention.

DEPOSIT INFORMATION

A deposit of the celery cultivar seed of this invention is maintained byA. Duda & Sons, Inc., 1200 Duda Trail, Oviedo, Florida 32765, U.S.A.Access to this deposit will be available during the pendency of thisapplication to persons determined by the Commissioner of Patents andTrademarks to be entitled thereto under 37 CFR § 1.14 and 35 USC § 122.Upon allowance of any claims in this application, all restrictions onthe availability to the public of the variety will be irrevocablyremoved by affording access to a deposit of at least 625 seeds of thesame variety with the American Type Culture Collection (ATCC), 10801University Boulevard, Manassas, Va. 20110 or National Collections ofIndustrial, Food and Marine Bacteria (NCIMB), 23 St Machar Drive,Aberdeen, Scotland, AB24 3RY, United Kingdom.

While a number of exemplary aspects and embodiments have been discussedabove, those of skill in the art will recognize certain modifications,permutations, additions, and sub-combinations thereof. It is thereforeintended that the following appended claims and claims hereafterintroduced are interpreted to include all such modifications,permutations, additions, and sub-combinations as are within their truespirit and scope.

What is claimed is:
 1. A seed, plant, or a plant part thereof, of celerycultivar designated TBG 45, wherein a representative sample of seed ofsaid cultivar has been deposited under ATCC Accession No. PTA-______. 2.A celery plant, or a plant part thereof, having essentially all of thephysiological and morphological characteristics of the celery plant ofclaim
 1. 3. A tissue or cell culture produced from protoplasts or cellsfrom the plant of claim 1, wherein said cells or protoplasts areproduced from a plant part selected from the group consisting of leaf,callus, pollen, ovule, embryo, cotyledon, hypocotyl, meristematic cell,root, root tip, pistil, anther, flower, seed, shoot, stem, stalk,petiole and sucker.
 4. A celery plant regenerated from the tissueculture of claim 3, wherein said regenerated plant comprises all of themorphological and physiological characteristics of celery cultivar TBG45.
 5. A method of producing a celery seed, wherein the method comprisescrossing the plant of claim 1 with a different celery plant andharvesting the resultant celery seed.
 6. A celery seed produced by themethod of claim
 5. 7. A celery plant, or a plant part thereof, producedby growing said seed of claim
 6. 8. A method of producing an herbicideresistant celery plant, wherein said method comprises introducing a geneconferring herbicide resistance into the plant of claim
 1. 9. Aherbicide resistant celery plant produced by the method of claim 8,wherein the gene confers resistance to a herbicide selected from thegroup consisting of glyphosate, sulfonylurea, imidazolinone, dicamba,glufosinate, phenoxy proprionic acid, L-phosphinothricin, cyclohexone,cyclohexanedione, triazine, benzonitrile protoporphyrinogen oxidase(PPO)-inhibitor herbicides, auxin herbicides, and broxynil.
 10. A methodof producing a pest or insect resistant celery plant, wherein saidmethod comprises introducing a gene conferring pest or insect resistanceinto the celery plant of claim
 1. 11. A pest or insect resistant celeryplant produced by the method of claim
 10. 12. The celery plant of claim11, wherein the gene encodes a Bacillus thuringiensis (Bt) endotoxin.13. A method of producing a disease resistant celery plant, wherein saidmethod comprises introducing a gene into the celery plant of claim 1.14. A disease resistant celery plant produced by the method of claim 13.15. A method of producing a celery plant with modified fatty acidmetabolism or modified carbohydrate metabolism comprising transformingthe celery plant of claim 1 with a transgene encoding a protein selectedfrom the group consisting of fructosyltransferase, levansucrase,α-amylase, invertase and starch branching enzyme or DNA encoding anantisense of stearyl-ACP desaturase.
 16. A celery plant having modifiedfatty acid metabolism or modified carbohydrate metabolism produced bythe method of claim
 15. 17. A method for producing a male sterile celeryplant, wherein said method comprises transforming the celery plant ofclaim 1 with a nucleic acid molecule that confers male sterility.
 18. Amale sterile celery plant produced by the method of claim
 17. 19. Amethod of introducing a desired trait into celery cultivar TBG 45,wherein the method comprises: (a) crossing a TBG 45 plant, wherein arepresentative sample of seed was deposited under ATCC Accession No.PTA-______, with a plant of another celery cultivar that comprises adesired trait to produce progeny plants, wherein the desired trait isselected from the group consisting of improved nutritional quality,industrial usage, male sterility, herbicide resistance, insectresistance, modified seed yield, modified lodging resistance, modifiediron-deficiency chlorosis and resistance to bacterial disease, fungaldisease or viral disease; (b) selecting one or more progeny plants thathave the desired trait to produce selected progeny plants; (c)backcrossing the selected progeny plants with the TBG 45 plants toproduce backcross progeny plants; (d) selecting for backcross progenyplants that have the desired trait; and (e) repeating steps (c) and (d)two or more times in succession to produce selected third or higherbackcross progeny plants that comprise the desired trait.
 20. A celeryplant produced by the method of claim 19, wherein the plant has thedesired trait and otherwise all of the physiological and morphologicalcharacteristics of celery cultivar TBG
 45. 21. The celery plant of claim20, wherein the desired trait is herbicide resistance and the resistanceis conferred to a herbicide selected from the group consisting ofimidazolinone, dicamba, cyclohexanedione, sulfonylurea, glyphosate,glufosinate, phenoxy proprionic acid, L-phosphinothricin, triazine,benzonitrile protoporphyrinogen oxidase (PPO)-inhibitor herbicides,auxin herbicides, and broxynil.
 22. The celery plant of claim 20,wherein the desired trait is insect resistance and the insect resistanceis conferred by a gene encoding a Bacillus thuringiensis endotoxin. 23.The celery plant of claim 20, wherein the desired trait is malesterility and the trait is conferred by a cytoplasmic nucleic acidmolecule.
 24. A method of producing a genetically modified celery plant,wherein the method comprises mutation, transformation, gene conversion,genome editing, RNA interference or gene silencing of the plant ofclaim
 1. 25. A genetically modified celery plant produced by the methodof claim 24, wherein the plant comprises the genetic modification andotherwise comprises all of the physiological and morphologicalcharacteristics of celery cultivar TBG 45.